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OBJECTIVES: To explore the postoperative kinesophobia of patients after percutaneous coronary intervention (PCI) and its related factors. BACKGROUND: Percutaneous coronary intervention is an effective method to treat coronary heart disease (CHD), and cardiac rehabilitation is an important auxiliary method after PCI. However, the compliance of patients with cardiac rehabilitation after PCI is not good, among which kinesophobia is an important influencing factor. DESIGN: A descriptive cross-sectional design was implemented, and the high-quality reporting of the study adhered to the Strengthening the Reporting of Observational Studies in Epidemiology Statement. METHODS: In total, 351 inpatients who underwent PCI in three tertiary grade-A hospitals in China were selected by convenient sampling method. We use one-way ANOVA and multiple linear regression analysis to determine the relevant related factors. RESULTS: The kinesophobia of patients after PCI was negatively correlated with chronic illness resource utilization and sense of personal mastery, and positively correlated with illness perception. Education level, clinical classification of CHD, exercise habits, chronic illness resource utilization, illness perception and sense of personal mastery entered the regression equation, which could explain 78.1% of the total variation. CONCLUSION: The level of kinesiophobia of patients after PCI is high. Education level, clinical classification of CHD, exercise habits, chronic illness resource utilization, illness perception and sense of personal mastery are the related factors of kinesiophobia of patients after PCI. RELEVANCE TO CLINICAL PRACTICE: By reducing the level of exercise fear of patients after PCI, patients are more likely to accept and adhere to the cardiac rehabilitation plan, thus improving their prognosis and improving their quality of life. PATIENT OR PUBLIC CONTRIBUTION: The patient underwent PCI in the research hospital. Researchers screen them according to the inclusion criteria and invite them to participate in this study. If they meet the requirements, participants will answer the research questionnaire face to face after signing the informed consent form.
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A high vascular patency was realized in the bulk or surface heparinized small-diameter in situ tissue-engineered vascular grafts (TEVGs) via a rabbit carotid artery replacement model in our previous studies. Those surface heparinized TEVGs could reduce the occurrence of aneurysms, but with a low level of the remodeled elastin, whereas those bulk heparinized TEVGs displayed a faster degradation and an increasing occurrence of aneurysms, but with a high level of the regenerated elastin. To combine the advantages of the bulk and surface graft heparinization to boost the remodeling of elastin and defer the occurrence of aneurysms, a coaxial electro-spinning technique was used to fabricate a kind of small-diameter core/shell fibrous structural in situ TEVGs with a faster degradable poly(lactic-co-glycolic acid) (PLGA) as a core layer and a relatively lower degradable poly(ε-caprolactone) (PCL) as a shell layer followed by the surface heparinization. The in vitro mechanical performance and enzymatic degradation tests revealed the resulting PLGA@PCL-Hep in situ TEVGs possessing not only a faster degradation rate, but also the mechanical properties comparable to those of human saphenous veins. After implanted in the rat abdominal aorta for 12 months, the good endothelialization, low inflammation, and no calcification were evidenced. Furthermore, the neointima layer of regenerated new blood vessels was basically constructed with a well-organized arrangement of elastin and collagen proteins. The results showed the great potential of these in situ TEVGs to be used as a novel type of long-term small-diameter vascular grafts.
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Cardiovascular disease has become one of the main causes of death. It is the common goal of researchers worldwide to develop small-diameter vascular grafts to meet clinical needs. Collagen is a valuable biomaterial that has been used in the preparation of vascular grafts and has shown good results. Recombinant humanized collagen (RHC) has the advantages of clear chemical structure, batch stability, no virus hazard and low immunogenicity compared with animal-derived collagen, which can be developed as vascular materials. In this study, Poly (l-lactide- ε-caprolactone) with l-lactide/ε-caprolactone (PLCL) and type III recombinant humanized collagen (hCOLIII) were selected as raw materials to prepare vascular grafts, which were prepared by the same-nozzle electrospinning apparatus. Meanwhile, procyanidin (PC), a plant polyphenol, was used to cross-link the vascular grafts. The physicochemical properties and biocompatibility of the fabricated vascular grafts were investigated by comparing with glutaraldehyde (GA) crosslinked vascular grafts and pure PLCL grafts. Finally, the performance of PC cross-linked PLCL-hCOLIII vascular grafts were evaluated by rabbit carotid artery transplantation model. The results indicate that the artificial vascular grafts have good cell compatibility, blood compatibility, and anti-calcification performance, and can remain unobstructed after 30 days carotid artery transplantation in rabbits. The grafts also showed inhibitory effects on the proliferation of SMCs and intimal hyperplasia, demonstrating its excellent performance as small diameter vascular grafts.
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The conversion of solar energy and water to hydrogen via semiconductor photocatalysts is one of the efficient strategies to mitigate the energy and environmental crisis. Conjugated polymeric photocatalysts have advantages over their inorganic counterparts. Their molecular structures, band structures, and electronic properties are easily tunable through molecular engineering to extend their spectral response ranges, improve their quantum efficiencies, and enhance their hydrogen evolution rates. In particular, covalent triazine-based frameworks (CTFs) present a strong potential for solar-driven hydrogen generation due to their large continuous π-conjugated structure, high thermal and chemical stability, and efficient charge transfer and separation capability. Herein, synthesis strategies, functional optimization, and applications in the photocatalytic hydrogen evolution of CTFs since the first investigation are reviewed. Finally, the challenges of hydrogen generation for CTFs are summarized, and the direction of material modifications is proposed.
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Increasingly growing problems in vascular access for long-term hemodialysis lead to a considerable demand for synthetic small diameter vascular prostheses, which usually suffer from some drawbacks and are associated to high failure rates. Incorporating the concept of in situ tissue engineering (TE) into synthetic small diameter blood vessels, for example, thermoplastic poly(ether urethane) (TPU) ones, could provide an alternative approach for vascular access that profits from the advantages of excellent mechanical properties of synthetic polymer materials (early cannulation) and unique biointegration regeneration of autologous neovascular tissues (long-term fistulae). In this study, a kind of heparinized small diameter (d = 2.5 mm) TPU/poly(ε-caprolactone) (TPU/PCL-Hep) bi-layered blood vessels was electrospun with an inner layer of PCL and an outer layer of TPU. Afterward, the inner surface heparinization was conducted by coupling H2N-PEG-NH2 to the corroded PCL layer and then heparin to the attached H2N-PEG-NH2 via the EDCI/NHS chemistry. Herein a heparinized PCL inner layer could not only inhibit thrombosis, but also provide sufficient space for the neotissue regeneration via biodegradation with time. Meanwhile, a TPU outer layer could confer the vascular access the good mechanical properties, such as flexibility, viability and fitness of elasticity between the grafts and host blood vessels as evidenced by the adequate mechanical properties, such as compliance (4.43 ± 0.07%/ 100 mmHg), burst pressure (1447 ± 127 mmHg) and suture retention strength (1.26 ± 0.07 N) without blood seepage after implantation. Furthermore, a rabbit carotid aortic replacement model for 5 months was demonstrated 100% animal survival and 86% graft patency. Puncture assay also revealed the puncture resistance and self-sealing (hemostatic time < 2 min). Histological analysis highlighted neotissue regeneration, host cell infiltration and graft remodeling in terms of extracellular matrix turnover. Altogether, these results showed promising aspects of small diameter TPU/PCL-Hep bi-layered grafts for hemodialytic vascular access applications.
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Substitutos Sanguíneos , Alicerces Teciduais , Animais , Artérias Carótidas/cirurgia , Poliésteres/química , Coelhos , Engenharia Tecidual/métodos , Alicerces Teciduais/químicaRESUMO
As thermoplastic polyurethane (TPU) elastomers possess good biocompatibility and mechanical properties similar to those of native vascular tissues, they were intended to be co-electrospun with poly(ε-caprolactone) (PCL) onto the outer surface of PCL electrospun small-diameter single-layer vascular grafts (SLVGs) in this study, combining with surface heparinization. In this work, a kind of structurally reinforced TPU+PCL/PCL small-diameter bilayer vascular graft (BLVG) was fabricated via layer-by-layer electrospinning followed by the heparinization of PCL via EDC/NHS chemistry. The resulting (TPU+PCL/PCL)-Hep BLVGs presented excellent mechanical strength and higher compliance, and sustainably released heparin exhibited enhanced anti-coagulation activity. During 6-month implantation in 18 rat abdominal aortas, these vascular prostheses induced the remodeling and regeneration of neovascular tissues, and promoted ECM deposition. Compared to heparinized PCL (PCL-Hep) SLVGs, the formation of aneurysm was completely inhibited and the onset of calcification was significantly delayed in (TPU+PCL/PCL)-Hep BLVGs. Not only vascular cell makers co-expressed by CD206+ cells were identified, but also a high content of elastin was evidenced due to the improvement of mechanical strength and compliance. These results indicated the feasibility and efficacy of inhibiting the aneurysm formation and boosting the vascular remodeling by incorporating TPU into PCL-Hep small-diameter artificial vascular grafts.
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Prótese Vascular , Poliuretanos , Animais , Aorta Abdominal/cirurgia , Poliésteres/química , Poliuretanos/química , Ratos , Alicerces Teciduais/químicaRESUMO
In order to reveal the mechanism of silicon (Si) fertilizer in improving nitrogen (N) and phosphorus (P) nutrient availability in paddy soil, we designed a series of soil culture experiments by combining application of varying Si fertilizer concentrations with fixed N and P fertilizer concentrations. Following the recommendations of fertilizer manufacturers and local farmers, we applied Si in concentrations of 0, 5.2, 10.4, 15.6, and 20.8 µg/kg. At each concentration of added Si, the availability of soil N and P nutrients, soil microbial activity, numbers of ammonia-oxidizing bacteria and P-decomposing bacteria which means that the organic P is decomposed into inorganic nutrients which can be absorbed and utilized by plants, and urease and phosphatase activity first increased, and then decreased, as Si was added to the soil. These indicators reached their highest levels with a Si application rate of 15.6 µg/kg, showing values respectively 19.78%, 105.09%, 8.34%, 73.12%, 130.36%, 28.12%, and 20.15% higher than those of the controls. Appropriate Si application (10.4 to 15.6 µg/kg) could significantly increase the richness of the soil microbial community involved in cycling of N and P nutrients in the soil. When the Si application rate was 15.6 µg/kg, parameters for characterizing microbial abundance such as sequence numbers, operational taxonomic unit (OTU) number, and correlation indices of microbial community richness such as Chao1 index, the adaptive coherence estimator (ACE) index, Shannon index, and Simpson index all reached maximum values, with amounts increased by 14.46%, 10.01%, 23.80%, 30.54%, 0.18%, and 2.64%, respectively, compared with the control group. There is also a good correlation between N and P mineralization and addition of Si fertilizer. The correlation coefficients between the ratio of available P/total P (AP/TP) and the number of ammonia-oxidizing bacteria, AP/TP and acid phosphatase activity (AcPA), AP/TP and the Shannon index, the ratio of available N/total amount of N (AN/TN) and the number of ammoniated bacteria, and AN/TN and AcPA were 0.9290, 0.9508, 0.9202, 0.9140, and 0.9366, respectively. In summary, these results revealed that enhancement of soil microbial community structure diversity and soil microbial activity by appropriate application of Si is the key ecological mechanism by which application of Si fertilizer improves N and P nutrient availability.
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Agricultura/métodos , Fertilizantes , Nitrogênio/química , Fósforo/química , Silício/química , Solo/química , Bactérias , Biodiversidade , Perfilação da Expressão Gênica , Microbiota , Nutrientes , Oryza , Análise de Componente Principal , Microbiologia do SoloRESUMO
Rapid endothelialization is crucial for in situ tissue engineering vascular grafts to prevent graft failure in the long-term. Gelatin is a promising nature material that can promote endothelial cells (ECs) adhesion, proliferation, and migration. In this study, the internal surface of electrospun polycaprolactone (PCL) vascular grafts was coated with gelatin. Endothelialization and vascular wall remolding were investigated by imaging and histological studies in the rat abdominal aorta replacement model. The endothelialization of heparinized gelatin-coated PCL (GP-H) vascular grafts was more rapid and complete than heparinized PCL (P-H) grafts. Intimal hyperplasia was milder in the GP-H vascular grafts than the P-H vascular grafts in the long-term. Meanwhile, smooth muscle cells (SMCs) and extracellular matrix (ECM) regeneration were better in the GP-H vascular grafts. By comparison, an aneurysm was observed in the P-H group in 6 months. Calcification was observed in both groups. All vascular grafts were patient after implantation in both groups. Our results showed that gelatin coating on the internal surface of PCL grafts is a simple and effective way to promote endothelialization. A more rapid endothelialization and complete endothelium can inhibit intimal hyperplasia in the long-term.
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Células Endoteliais , Gelatina , Animais , Prótese Vascular , Humanos , Poliésteres , RatosRESUMO
Decellularization is a promising approach in tissue engineering to generate small-diameter blood vessels. However, some challenges still exist. We performed two decellularization phases to develop an optimal decellularized scaffold and analyze the relationship between the extracellular matrix (ECM) composition and mechanical properties. In decellularization phase I, we tested sodium dodecylsulfate (SDS), Triton X-100 (TX100) and trypsin at different concentrations and exposure times. In decellularization phase II, we systematically compared five combined decellularization protocols based on the results of phase I to identify the optimal method. These protocols tested cell removal, ECM preservation, mechanical properties, and residual cytotoxicity. We further immobilized heparin to optimal decellularized scaffolds and determined its anticoagulant activity and mechanical properties. The combined decellularization protocol comprising treatment with 0.5% SDS followed by 1% TX100 could completely remove the cellular contents and preserve the mechanical properties and ECM architecture better. In addition, the heparinized decellularized scaffolds not only had sustained anticoagulant activity, but also similar mechanical properties to native vessels. In conclusion, heparinized decellularized scaffolds represent a promising direction for small-diameter vascular grafts, although further in vivo studies are needed.
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Heparina/metabolismo , Fenômenos Mecânicos , Alicerces Teciduais/química , Animais , Matriz Extracelular/metabolismo , Octoxinol/química , Dodecilsulfato de Sódio/química , Tripsina/químicaRESUMO
This study was conducted to reveal the effects of silicon (Si) application on nutrient utilization efficiency by rice and on soil nutrient availability and soil microorganisms in a hybrid rice double-cropping planting system. A series of field experiments were conducted during 2017 and 2018. The results showed that Si nutrient supply improved grain yield and the utilization rates of nitrogen (N) and phosphorus (P) to an appropriate level for both early and late plantings, reaching a maximum at 23.4 kg/ha Si. The same trends were found for the ratios of available N (AN) to total N (TN) and available P (AP) to total P (TP), the soil microbial biomass carbon (MBC), microbial biomass nitrogen (MBN), microbial biomass phosphorus (MBP), and the ratios of MBN to TN and MBP to TP, at different levels of Si. Statistical analysis further revealed that Si application enhanced rice growth and increased the utilization rate of fertilizer due to an ecological mechanism, i.e., Si supply significantly increased the total amount of soil microorganisms in paddy soil compared to the control. This promoted the mineralization of soil nutrients and improved the availability and reserves of easily mineralized organic nutrients.
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Nitrogênio/metabolismo , Oryza/crescimento & desenvolvimento , Fósforo/metabolismo , Silício/metabolismo , Microbiologia do Solo , Agricultura/métodos , Biomassa , Carbono , Fertilizantes , Nutrientes/metabolismo , Solo/químicaRESUMO
To determine whether the newly built lead-acid battery agglomeration area in a town in northern Zhejiang had an impact on the surrounding soil environment after seven years of operation, 76 samples of surface soil around the lead-acid battery concentration area were collected, and the contents of Hg, As, Cu, Zn, Pb, Cd, Ni, and Cr in the soil were determined. Based on the spatial distance of the agglomeration area and 50, 450, and 850 m from the agglomeration area boundary, the soil environmental quality was evaluated using the single factor index, Nemerow comprehensive pollution index, and potential ecological risk index methods. The spatial distribution characteristics of the heavy metals were analyzed using the geostatistical method, and the sources of heavy metals affecting the soil environment were determined by correlation analysis. The results showed that the average contents of Hg, Zn, and Pb in eight heavy metals were higher than their corresponding background values at all spatial scales. The average Cd content in spatial scales other than the agglomeration area was larger than its background value. Only As at 50 m was greater than its background value, whereas the average content of other elements at all spatial distances was lower than their corresponding background values. The spatial variability was high for Hg and Cd but not obvious for other elements. This implies that the influence of regional activities was concentrated mainly on Hg and Cd; the content of both increased with distance from the agglomeration area. Hg and Cd exceeded the risk screening values and were distributed mainly at 450 m and 850 m; 33.33% and 38.89% Hg points and 27.78% and 55.56% Cd points were observed at these distances, respectively. The spatial distribution characteristics of Hg and Cd were consistent with their contents; only Zn and Pb had scattered points that exceeded the risk screening values and generally no obvious spatial distribution characteristics. According to the risk analysis of soil comprehensive pollution caused by the eight heavy metals, Cd was the main source of soil comprehensive pollution risk at a contribution rate of 36.73%, which caused the soil at 850 m to be in a state of alert. Soil ecological risk at a medium level occurred mainly at 450 m and 850 m outside the agglomeration area from Hg and Cd. The contribution rates to the soil quality at these distances were 46.30% and 39.37% for Hg and 38.98% and 49.30% for Cd, respectively. This indicates that regional activities caused Hg and Cd to be the main elements affecting soil quality in the study area. The results of geostatistics and multivariate statistical analysis showed that Hg and Cd were diffused inward from the periphery of the agglomeration area on the axis of the local main wind direction (northeast-southwest), and the main sources of both were coal-burning activities of enterprises in the periphery of the agglomeration area. In summary, the newly build lead-acid battery agglomeration area has not significantly affected the accumulation of heavy metals in the agglomeration area and in the surrounding soil after seven years of operation.
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The study investigated the potential of ensuring safe production of rice from paddy fields affected by heavy cadmium contamination. A paddy soil polluted with heavy cadmium (total Cd content of 2.52 mg·kg-1) surrounding a lead-zinc mining area in Guiyang County of Chenzhou City, Hunan province was selected for analysis. We investigated production using a low cadmium accumulation rice variety (Oryza sativa L. Jia-33) and passivation additives for heavy metal activity (including humic acid and sepiolite). Results showed that:Oryza sativa L. Jia-33 showed good low cadmium accumulation characteristics in rice fields with heavy cadmium pollution. When humic acid and sepiolite were applied (alone or combined), the accumulation of Cd in different organs of the rice declined, and the transport coefficient of Cd for stem to root and polished rice to stem also declined. The rate of decline increased with increased application dosage. The cadmium content in polished rice was lower than the limit established in the National Food Safety Standard (GB 2715-2012), when applying 5.250 t·hm-2 humic acid, 6.750 t·hm-2 sepiolite, or a combination of 1.125 t·hm-2 humic acid and 3.37 5 t·hm-2 sepiolite; Cd content in polished rice was (0.171±0.01), (0.184±0.01), and (0.181±0.01) mg·kg-1, respectively. Single or combined application of humic acid and sepiolite promoted the transformation of soil Cd to residual and Fe Mn oxide bound forms, significantly reducing the content of available Cd and the bioavailability of Cd in soil, and further reducing Cd content in all rice organs. The reduction was more effective with single application of sepiolite or combined application of humic acid and sepiolite. The effects of combined application of humic acid and sepiolite were also more beneficial for soil nutrients, while soil available phosphorus, available potassium, phosphorus, and organic matter content increased with increased application of amendments. Soil nitrogen content did not change. In conclusion, results indicate that joint application of humic acid, sepiolite, and a low Cd accumulation rice variety are best for safe production on heavy cadmium-contaminated soil.
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Cádmio/análise , Substâncias Húmicas/análise , Silicatos de Magnésio/análise , Oryza , Poluentes do Solo/análise , China , Mineração , SoloRESUMO
In order to research the coordination relation between sepiolite application and the contents of available lead(Pb) and cadmium(Cd) in soil or rice absorption and redistribution of Pb and Cd, a pot experiment was conducted by selecting the soil sampled from a area polluted by Pb and Cd combination in Shaoxing City of Zhejiang Province and atypical late Japonica Rice Varieties in Zhejiang Province(Jia 33) with lower accumulation for Pb, Cd. The results showed that:the contents of available Pb and Cd in soil were significantly negatively correlated with the concentration of sepiolite added, the correlation coefficients of contents of available Pb and Cd with the concentration of sepiolite added were -0.940 respectively and -0.952, which reached the significant level(P<0.01).The contents of Pb and Cd in rice roots, stems, leaves and polished rice had different reducing rates with the increase of the concentration of sepiolite. In addition, with the increase of the concentration of sepiolite added, the enrichment coefficients of Pb and Cd in rice roots, stems, leaves and polished rice decreased dramatically. At the same time, Pb and Cd absorbed by roots transferred to the stems decreasingly, Pb and Cd absorbed by stems transferred to the polished rice decreasingly. When the addition of Sepiolite was 9.00 g·kg-1 soil, the contents of Pb and Cd in polished rice of Jia 33 were(0.14±0.02) mg·kg-1 and(0.03±0.01) mg·kg-1 respectively, which were all lower than the national limit index(GB 2762-2012). Compared with the control group, the enrichment coefficients of Pb in roots, stems and leaves and polished rice decreased by 8.83%, 29.96%, 49.20%, 79.41% respectively, and the enrichment coefficients of Cd decreased by 23.08%, 63.22%, 44.00%, 82.35% respectively. In addition, the transportation coefficients of Pb and Cd absorbed by roots transfer to the stems were decreased by 23.18% and 52.19% respectively and the transportation coefficients of stem rice translocation of Pb and Cd absorbed by stems transfer to the polished rice were decreased by 70.83%, 52.00%, respectively. This result means the sepiolite had a good resistance control in the migration and redistribution of Pb and Cd in soil rice system on the soil polluted by lead and cadmium, indicating the combination of rational application of sepiolite and rice varieties with lower accumulation for heavy metals can achieve safe agricultural utilization of the soil with combined contamination by a relatively higher concentration of Pb and Cd.
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Cádmio/análise , Chumbo/análise , Silicatos de Magnésio/química , Oryza/química , Poluentes do Solo/análise , Solo/químicaRESUMO
We herein report a water-stable 3D dysprosium-based metal-organic framework (MOF) that can non-covalently interact with probe ss-DNA. The formed system can serve as an effective fluorescence sensing platform for the detection of complementary Ebolavirus RNA sequences with the detection limit of 160 pM.
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Disprósio/química , Ebolavirus/isolamento & purificação , Corantes Fluorescentes/química , Doença pelo Vírus Ebola/diagnóstico , Compostos Organometálicos/química , RNA Viral/análise , Sequência de Bases , Cristalografia por Raios X , Sondas de DNA/química , Sondas de DNA/genética , Ebolavirus/genética , Doença pelo Vírus Ebola/virologia , Humanos , Limite de Detecção , Modelos Moleculares , RNA Viral/genética , Espectrometria de Fluorescência/métodos , Água/químicaRESUMO
OBJECTIVE: To investigate the active efflux of fluoroquinolones in clinical isolated strains of Escherichia coli. METHODS: The accumulation of ciprofloxacin and tosufloxacin for clinical isolates of Escherichia coli and the effect of glucose and Carbonyl cyanide mchlorophenyl hydrazone (CCCP) on the uptake of Escherichia coli were determined by a modified fluorometric assay. RESULTS: The accumulation of ciprofloxacin in sensitive and multi-drug-resistant (MDR) organisms decreased with energy dependence, and the decrease in MDR isolate was prominent. Incubation with CCCP increased the accumulation of ciprofloxacin up to three-fold in MDR isolate but not in sensitive isolate. In contrast, CCCP appeared to have no influence on the uptake of tosufloxacin in Escherichia coli. CONCLUSION: These results indicate that energy-dependent active efflux is one of the mechanisms of hydrophilic fluoroquinolones resistance, but it does not play any role in hydrophobic fluoroquinolones resistance in Escherichia coli.
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Anti-Infecciosos/metabolismo , Escherichia coli/efeitos dos fármacos , Fluoroquinolonas/farmacologia , Anti-Infecciosos/farmacologia , Farmacorresistência Bacteriana Múltipla , Escherichia coli/isolamento & purificação , Escherichia coli/metabolismo , Humanos , Testes de Sensibilidade MicrobianaRESUMO
PURPOSE: The ETS family of transcription factors regulate several critical cellular functions. They have also been implicated in invertebrate ocular development. This work was undertaken to determine whether epithelium-specific ETS transcription factors are expressed in the retinal pigment epithelium and to investigate the possible role of these factors in retinal diseases such as age-related macular degeneration. METHODS: The expression of the epithelial ETS transcription factors ELF5, ESE3, and ELF3 was assessed by RT-PCR in the human RPE cell lines D407 and hTERT-RPE1. The full-length coding sequence of rat Elf3 was isolated with 3' rapid amplification of cDNA ends (RACE) and degenerative primers, and its expression was determined in various rat tissues, by RT-PCR and real-time PCR. A polyclonal ELF3 antibody produced from a C-terminal peptide was used to observe the distribution of the transcription factor within the retina. To assess the possible ELF3 regulation of the TIMP3 promoter, transient transfection assays were performed. Promoter activity was determined with a firefly luciferase reporter gene construct. RESULTS: The epithelium-specific ETS transcription factor ELF3 was expressed in the D407 and hTERT-RPE1 cell lines. Neither ESE3 nor ELF5 was detected in the RPE. The cloning of rat Elf3 produced two splice variants, designated Elf3a (1786 bp) and Elf3b (1855 bp). The larger form, Elf3b, contained a 69-bp insert in the coding sequence, which showed high homology to a similar insert previously identified in murine Elf3. Both splice variants were expressed in rat lung, kidney, liver, and retina, but were absent in heart tissue. Real-time PCR analysis showed the retina to contain high levels of Elf3, which was subsequently localized to the RPE. Elf3 upregulated the TIMP3 promoter, with Elf3a and -3b inducing an approximate sixfold increase in activity. CONCLUSIONS: The ELF3 transcription factor is highly expressed in the RPE and can regulate important ocular genes, such as TIMP3, in vitro. The specific expression of ELF3 in the RPE may reflect an important role for this transcription factor in retinal function. Furthermore, its regulation of TIMP3 may have implications for degenerative retinal diseases, such as age-related macular degeneration.
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Proteínas de Ligação a DNA/genética , Proteínas do Olho/genética , Epitélio Pigmentado Ocular/metabolismo , Proteínas Proto-Oncogênicas/genética , Retina/metabolismo , Fatores de Transcrição/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Western Blotting , Linhagem Celular , Proteínas de Ligação a DNA/metabolismo , Proteínas do Olho/metabolismo , Humanos , Dados de Sequência Molecular , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Proto-Oncogênicas c-ets , Coelhos , Ratos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Distribuição Tecidual , Inibidor Tecidual de Metaloproteinase-3 , Inibidores Teciduais de Metaloproteinases/metabolismo , Fatores de Transcrição/metabolismoRESUMO
OBJECTIVE: To investigate the effects of Carthamus tinctorius on bcl-2, caspase-3 expression of apoptosis of neurons. METHOD: SD rats were randomly divided into ischemia control group, large-dose group, middle-dose group and low-dose group. The middle cerebral artery of rats was occluded for 2h by inserting an introluminal molofilament, and reperfusion was then instituted for 4h or 22h. The brains were stained with 2, 3, 5-triphenylte trazolinm chloride for assessment of volume of infarction, and then embedded onto slides with paraffin for morphological assessment and immunohistochemistry was carried out to investigate the changes in bcl-2 and caspase-3. RESULT: All treated groups at different times decreased the volume of infarction (P < 0.05), while large-dose group showed more distinct decrease than other groups (P < 0.05). All treated groups at different times increased bcl-2 and decreased caspase-3 expression as well, while large-dose group showed more distinct effect (P < 0.05). CONCLUSION: C. tinctorius injection can reduce the volume of cerebral infarction, and increased bcl-2 and decreased caspase-3 expression.
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Encéfalo/patologia , Carthamus tinctorius , Caspases/metabolismo , Infarto da Artéria Cerebral Média/patologia , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Animais , Apoptose/efeitos dos fármacos , Encéfalo/metabolismo , Carthamus tinctorius/química , Caspase 3 , Relação Dose-Resposta a Droga , Medicamentos de Ervas Chinesas/administração & dosagem , Medicamentos de Ervas Chinesas/farmacologia , Feminino , Infarto da Artéria Cerebral Média/metabolismo , Masculino , Neurônios/citologia , Neurônios/metabolismo , Plantas Medicinais/química , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Traumatismo por Reperfusão/metabolismo , Traumatismo por Reperfusão/patologiaRESUMO
AIM:To study the cell types, localization, distribution density and morphology of APUD cells in the intestinal mucosa of stomachless teleost fishes.METHOD:By using the peroxidase antiperoxidase complex (PAP) immunocytochemical staining technique the identification, localization and morphology of immunoreactive (IR) endocrine cells seattered in the intestinal mucosa of grass carp (Cyenopharyngodon idellus), black carp ( Mylopharyngodon piceus ) and common carp (Cyprinus carpio) were investigated with 20 kinds of antisera prepared against mammalian peptide hormones of APUD cells, and likewise by using avidin-biotin-peroxidase complex (ABC) method those of silver carp (Hypophthalmichthys molitrix), bighead (Aristichthys nobilis), silver crucian carp (Carassius gibelio) and bluntnose black bream (Megalobrama amblyocephala ) were also studied with 5 different antisera. The replacement of the first antiserum by phosphate buffered saline (PBS) was employed as a control. IR endocrine cells were counted with a square-mesh ocular micrometer from 10 fields selected randomly in every section of each part of the intestine specimen. The average number of IR endocrine cells per mm(2) was counted to quantify their distribution density.RESULT:Gastrin (GAS), Gastric inhibitory peptide (GIP), glucagon (GLU), glucagons like immunorea-ctants (GLI), bovine pancreatic polypeptide (BPP), leucine-enkephalin (ENK) and substance P (SP)-IR endocrine cells were found in the gut of grass carp, black carp and common carp, and somatostatin (SOM) IR endocrine cells were only seen in common carp. GAS, GIP and GLU-IR endocrine cells were found in the intestinal mucosa of silver carp, bighead, silver crucian carp and bluntnose black bream. Most of IR endocrine cells had the higher distribution density in the foregut and midgut, and were longer in shape. They had a long apical cytoplasmic process extended to the gut lumen and a basal process extended to adjacent cells or basement membrane and touched with it. Sometimes, the basal cytoplasmic process formed an enlarged synapse-like structure in the contiguous part with basement membrane. This phenomenon provided new morpho-logical evidence for neuroendocrine and paracrine secretory function of these enteroendocrine cells.CONCLUTION:At least 8 kinds of IR endocrine cells were found in the gut of stomachless teleost species for the first time in China. These IR endocrine cells scattering in the gut mucosa belong to the APUD system. Among them, the hormones secreted by SP-, ENK-, SOM- and GLU-IR endocrine cells belong to the peptides of dual distribution in the brain and gut. This provided new evidence for the concept of brain-gut peptide. According to the cell types, distribution density, morphological characteristics and variety in shape of APUD cells in the gut of stomachless teleost fishes, it is deemed that the digestive tract of fishes is also an endocrine organ of great importance and complexity.
RESUMO
AIM:To identify the type localization and morphology of APUD endocrine cells in the gastroenteropancreatic (GEP) system of stomach-containing teleosts, and study APUD endocrine system in the stomach, intestine and pancreas of fish species.METHODS:Two kinds of immunocytochemical (ICC) techniques of the streptavidin biotin-peroxidase complex (SABC) and streptavidin-peroxidase (S-P) method were used. The identification, localization and morphology of APUD endocrine cells scattered in the mucosa of digestive tract, intermuscular nerve plexus and glandular body of northern snakehead (Channa argus), ricefield eel (Monopterus albus), yellow catfish (Pelteobagrus fulvidraco), mandarinfish (Siniperca chuatsi), largemouth bass (Micropterus salmoides),oriental sheatfish (Silurus asotus), freshwater pomfret (Colossoma brachypomum) and nile tilapia (Tilapia nilotica) were investigated with 8 kinds of antisera.RESULTS:The positive reaction of 5-hydroxytryptamine (5-HT) immunoreactive endocrine (IRE) cells was found in the digestive tract and glandular body of 8 fish species in different degree.Only a few gastrin (GAS)-IRE cells were seen in C.argus,M.albusand P.fulvidraco. Glucagon (GLU)IRE cells were not found in the digestive tract and glandular body but existed in pancreatic island of most fish species. The positive reaction of growth hormone (GH)IRE cells was found only in pancreatic island of S. Chuatsi and S. Asotus, no positive reaction in the other 6 fish species. Somatostatin (SOM), calcitonin (CAL), neurofilament (NF) and insulin (INS)-IRE cells in the stomach, intestine and pancreas of 8 kinds of fish were different in distribution and types. The distribution of all 8 APUD cells was the most in gastrointestinal epithelium mucosa and then in digestive glands. The positive reaction of SOM and 5-HT-IRE cells was found in intermuscular nerve plexus of intestine of P.fulvidraco and S.chuatsi. Only GH-IRE cells were densely scattered in the pancreatic islands of S.chuatsi and S. asotus, and odd distribution in the pancreas of S. asotus.SOM-IRE cells were distributed in the pancreatic islands of S. asotus, C. Brachypomumand T. nilotica. There were INS-IRE cells in the pancreatic islands of S. chuatsi and S. asolus. Eight kinds of APUD cells had longer cell body and cytoplasmic process when they were located in the gastrointestinal epithelium, and had shorter cell body and cytoplasmic process in the gastric gland, and irregular shape in the esophagus and pancreatic island.CONCLUSION:Eight kinds of IRE cells were identified in the GEP system of stomach-containing teleosts. These endocrine cells were scattered in gastrointestinal mucosa, intermuscular nerve plexus, gland body, pancreatic gland and islands under APUD system. CAL and GH-IRE cells in the pancreatic islands of fishes showed functional diversity for these two hormones. Their morphological feature provides evidence of endocrine-paracrine and endocrine-exocrine acting mode. This research can morphologically prove that the GEP endocrine system of fish (the lowest vertebrate) is almost the same as of mammal and human.