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A molecularly imprinted ratiometric fluorescent sensor was synthesized for the detection of 4-bromophenoxybenzene (BDE-3) based on perovskite quantum dots and metal organic framework. First, aspartic acid (Asp) was introduced during the synthesis of perovskite CsPbX3 for the formation of a core-shell structure of CsPbX3@Asp-Cs4PbX6. Due to the protection of the shell layer Cs4PbX6, the stability of the core CsPbX3 was improved significantly. Compared to CsPb(BrI)3, the ultraviolet and thermal stabilities of CsPb(BrI)3@Asp-Cs4Pb(BrI)6 were increased by 26 times and 32 times, respectively, and, compared to CsPbBr3, these stabilities of CsPbBr3@Asp-Cs4PbBr6 were increased by 3 times and 13 times, respectively. The water stabilities of CsPb(BrI)3@Asp-Cs4Pb(BrI)6 and CsPbBr3@Asp-Cs4PbBr6 were greatly improved too. Then, a ratiometric fluorescence sensor was constructed by in situ growth of CsPb(BrI)3@Asp-Cs4Pb(BrI)6 in metal organic framework (NH2-MIL-53) for the detection of BDE-3, in which the orange fluorescence of CsPb(BrI)3@Asp-Cs4Pb(BrI)6 (614 nm) was regarded as the reference signal and the cyan fluorescence of NH2-MIL-53 (494 nm) was used as the fluorescence response signal. To improve the selectivity of the sensor, the molecular imprinting polymer (MIP) was modified on the NH2-MIL-53 and an imprinting factor of 3.17 was obtained. Under 365 nm light excitation, the fluorescent response signal at 494 nm was quenched gradually by BDE-3 in the range 11.4 to 68.5 nmol/L, while the reference signal at 614 nm remained unchanged. The limit of detection and limit of quantification were 3.35 and 11.2 nmol/L, respectively, and the fluorescent color of the sensor changed gradually from cyan to green to orange, which illustrated that the developed sensor has an ability to recognize BDE-3 specifically, a good anti-interference ability, and a sensitively visual detection ability. Moreover, the sensor was successfully applied to the BDE-3 detection in polyethylene terephthalate plastic bottle, polyvinyl chloride plastic bag, and circuit board with satisfactory recoveries (96.3-108.1%) and low relative standard deviations (5%). The preparation processes of NH2-MIL-53, NH2-MIL-53-CsPb(BrI)3@Asp-Cs4Pb(BrI)6, and the MIP-NH2-MIL-53-CsPb(BrI)3@Asp-Cs4Pb(BrI)6 composites.
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In this Letter, we investigate a snapshot spectral-polarimetric-volumetric imaging (SSPVI) system using a single detector. Through compressed acquisition and reconstruction, SSPVI can achieve spectral imaging (x,y,λ), polarization imaging (x,y,ψ,χ), and light field imaging (x,y,θ,φ) simultaneously. The newly discovered performance is showcased by attaining the spectral-polarimetric-volumetric video and different laboratory accuracy experiments. These never-seen-before capacities of the camera open new prospects for many applications, such as biological analysis, object recognition, and remote sensing.
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Background: To explore the predictive value of placental features in early pregnancy for gestational diabetes mellitus (GDM) using deep and radiomics-based machine learning (ML) applied to ultrasound imaging (USI), and to develop a nomogram in conjunction with clinical features. Methods: This retrospective multicenter study included 415 pregnant women at 11-13 weeks of gestation from two institutions: the discovery group from center 1 (n=305, control group n=166, GDM group n=139), and the independent validation cohort (n=110, control group n=57, GDM group n=53) from center 2. The 2D USI underwent pre-processed involving normalization and resampling. Subsequently, the study performed screening of radiomics features with Person correlation and mutual information methods. An RBF-SVM model based on radiomics features was constructed using the five-fold cross-validation method. Resnet-50 as the backbone network was employed to learn the region of interest and constructed a deep convolutional neural network (DLCNN) from scratch learning. Clinical variables were screened using one-way logistic regression, with P<0.05 being the threshold for statistical significance, and included in the construction of the clinical model. Nomogram was built based on ML model, DLCNN and clinical models. The performance of nomogram was assessed by calibration curves, area under the receiver operating characteristic curve (AUC) and decision curve analysis (DCA). Results: The AUCs for the ML model in the discovery cohort and independent validation cohort were 0.91 (0.88-0.94) and 0.86 (0.79-0.93), respectively. And 0.65 (0.59-0.71), 0.69 (0.59-0.79) for the DLCNN, 0.66 (0.59-0.72), 0.66 (0.55-0.76) for the clinical model, respectively. The nomogram exhibited the highest performance with AUCs of 0.93 (0.90-0.95) and 0.88 (0.81-0.94) The receiver operating characteristic curve (ROC) proved the superiority of the nomogram of clinical utility, and calibration curve showed the goodness of fit of the model. The DCA curve indicated that the nomogram outperformed other models in terms of net patient benefit. Conclusions: The study emphasized the intrinsic relationship between early pregnancy placental USI and the development of GDM. The use of nomogram holds potential for clinical applications in predicting the development of GDM.
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Inteligência Artificial , Diabetes Gestacional , Gravidez , Feminino , Humanos , Ultrassom , Diabetes Gestacional/diagnóstico , Placenta/diagnóstico por imagem , Redes Neurais de ComputaçãoRESUMO
Background: The effect of intravenous (IV) vitamin C in the treatment of sepsis remains controversial. We aimed to explore the clinical efficacy of vitamin C in the treatment of sepsis. Methods: Electronic databases (PubMed, Embase, Web of Science, and Cochrane Library) were searched from inception through November 15th, 2022, for randomized controlled trials evaluating the effect of IV vitamin C treatment in patients with sepsis. The primary outcome was short-term mortality, secondary outcomes included duration of vasopressor use, length of intensive care unit (ICU) stay, and Sequential Organ Failure Assessment (SOFA) score after vitamin C treatment. Subgroup analyses were performed based on the dose and duration of IV vitamin C and region to determine whether vitamin C benefited patients with sepsis. Results: A total of 10 studies including 1,426 patients fulfilled the predefined criteria and were analyzed. Overall, there were no significant differences between the vitamin C group and the control group regarding short-term mortality [odds ratio (OR), 0.61; 95% confidence interval (CI) 0.37-1.01; p = 0.05], ICU length of stay [mean difference (MD), -1.24; 95% CI -3.54 to 1.05, p = 0.29] and SOFA score (MD, -0.85, 95% CI -2.38 to 0.67, p = 0.27). However, vitamin C significantly reduced the duration of vasopressor use (MD, -14.36, 95% CI -26.11 to -2.61, p = 0.02). Furthermore, subgroup analysis found that in developing countries, vitamin C was associated with a significant reduction in short-term mortality (OR, 0.33; 95% CI 0.12-0.90; p = 0.03), duration of vasopressor use (MD, -24.37, 95% CI -33.72 to -15.02, p < 0.001) and SOFA score (MD, -2.55, 95% CI -4.81 to -0.28, p = 0.03). Conclusion: In our study, vitamin C administration for sepsis patients was not associated with a significant reduction in short-term mortality, length of ICU stay or SOFA score. However, we observed that vitamin C could reduce the duration of vasopressor use. Furthermore, sepsis patients in developing countries may benefit more from vitamin C administration than those in developed countries.Systematic review registration: Identifier CRD42022380958, https://www.crd.york.ac.uk/PROSPERO/display_record.php?RecordID=380958.
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Background: AKT3 appears to play a role in lung cancer. However, its role in ventilator-associated pneumonia is still unclear. Therefore, this study aimed to investigate the role of AKT3 in macrophages during ventilator-associated pneumonia. Methods: The mRNA level of AKT3, Data from The Cancer Genome Atlas (TCGA), Gene Expression Omnibus (GEO), The data is analyzed using the Xiantao academic analysis tool. Additionally, the roles of AKT3 in ventilator-associated pneumonia (VAP) were investigated through in vivo experiments. Results: AKT3 was differentially expressed in various normal and tumor tissues. Functional enrichment analysis indicated the immunomodulatory function and inflammatory response of AKT3 in lung cancer. Depletion of macrophages protected against lung epithelial cells and significantly decreased MMP9, MMP19, FTH, and FTL expression levels and increased GPX4 expression levels, while partially reversing the changes in macrophage. Mechanistically, macrophage depletion attenuates ferroptosis of lung epithelial cells by modulating AKT3 following VAP. Conclusion: Collectively, this study suggests the need for further validation of the immunoregulatory function of AKT3 in lung cancer. Additionally, macrophage depletion mitigates lung injury by modulating the AKT3/GPX4 pathway in the context of VAP.
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Lesão Pulmonar , Neoplasias Pulmonares , Pneumonia Associada à Ventilação Mecânica , Humanos , Células Epiteliais , Macrófagos , Proteínas Proto-Oncogênicas c-aktRESUMO
Identifying the key factors mediating the progression from hypertension to cardiac hypertrophy is critically important for developing a strategy to protect against heart failure. Serum exosomes have been revealed to be involved in the development of cardiovascular disease. In the current study, we found that either serum or serum exosomes derived from SHR induced hypertrophy in H9c2 cardiomyocytes. SHR Exo injection through the tail vein for 8 weeks induced left ventricular wall thickening and decreased cardiac function in C57BL/6 mice. SHR Exo carried the renin-angiotensin system (RAS) proteins AGT, renin, and ACE into cardiomyocytes, which increased the autocrine secretion of Ang II. Moreover, the AT1-type receptor antagonist telmisartan prevented hypertrophy of H9c2 cells induced by SHR Exo.These results identified a novel role of exosomes derived from SHR serum in cardiac hypertrophy and revealed that SHR Exo induced cardiac hypertrophy by carrying AGT, renin, and ACE proteins into cardiomyocytes to increase their autocrine secretion of Ang II. The emergence of this new mechanism will help us better understand how hypertension progresses to cardiac hypertrophy.
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Exossomos , Hipertensão , Ratos , Camundongos , Animais , Angiotensina II/metabolismo , Ratos Endogâmicos SHR , Miócitos Cardíacos/metabolismo , Renina/metabolismo , Exossomos/metabolismo , Camundongos Endogâmicos C57BL , Cardiomegalia/induzido quimicamente , Cardiomegalia/metabolismo , Sistema Renina-Angiotensina , Hipertensão/metabolismoRESUMO
This study was aimed to develop an efficient tumour-targeted liposome nanobubbles (LNBs) system using ultrasound-targeted nanobubble destruction for enhanced release and transfection of miRNA-199a-3p in hepatocellular carcinoma (HCC) therapy. The prepared LNBs comprised a polyethylene glycol-modified liposome shell and a perfluoropentane (PFP) core. MiRNA-199a-3p was attached to the nanocomposite surface via electrostatic adsorption, while RGD peptide functionalized the LNBs surface for enhanced HCC cell targeting, namely PFP@miR-RGD-LNBs. The LNBs were spherical with a narrow size distribution. The gene-loaded LNBs effectively condensed miR-199a-3p and protected it from enzymatic degradation. Low-intensity focused ultrasound (LIFU) promoted a fast release of miR-199a-3p from the prepared LNBs, thereby enhancing therapeutic effects. The combined application of PFP@miR-RGD-LNBs and LIFU exhibited a more potent inhibitory effect on HepG2 cells than the other groups, potentially due to LIFU promoting rapid and efficient gene release at the target site and increasing cell membrane permeability. Quantitative reverse transcription-polymerase chain reaction analysis revealed significantly increased mRNA expression levels of key apoptosis markers (Bad, Bax, Caspase-9 and Caspase-3) in the PFP@miR-RGD-LNBs + LIFU group compared to other groups. These findings suggest that the prepared LNBs are highly likely to be promising candidates for further exploration of HCC gene delivery and therapy.
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Carcinoma Hepatocelular , Neoplasias Hepáticas , MicroRNAs , Humanos , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/terapia , Carcinoma Hepatocelular/metabolismo , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/terapia , Neoplasias Hepáticas/metabolismo , Lipossomos , MicroRNAs/genética , MicroRNAs/metabolismo , Oligopeptídeos/metabolismo , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , Linhagem Celular TumoralRESUMO
Monocyte-derived exosomes (Exos) have been implicated in inflammation-related autoimmune/inflammatory diseases via transferring bioactive cargoes to recipient cells. The purpose of this study was to investigate the possible effect of monocyte-derived Exos on the initiation and the development of acute lung injury (ALI) by delivering long non-coding RNA XIST. Key factors and regulatory mechanisms in ALI were predicted by bioinformatics methods. BALB/c mice were treated with lipopolysaccharide (LPS) to establish an ALI in vivo model and then injected with Exos isolated from monocytes transduced with sh-XIST to evaluate the effect of monocyte-derived exosomal XIST on ALI. HBE1 cells were co-cultured with Exos isolated from monocytes transduced with sh-XIST for further exploration of its effect. Luciferase reporter, RIP and RNA pull-down assays were performed to verify the interaction between miR-448-5p and XIST, miR-448-5p and HMGB2. miR-448-5p was significantly poorly expressed while XIST and HMGB2 were highly expressed in the LPS-induced mouse model of ALI. Monocyte-derived Exos transferred XIST into HBE1 cells where XIST competitively inhibited miR-448-5p and reduced the binding of miR-448-5p to HMGB2, thus upregulating the expression of HMGB2. Furthermore, in vivo data revealed that XIST delivered by monocyte-derived Exos downregulated miR-448-5p expression and up-regulated HMGB2 expression, ultimately contributing to ALI in mice. Overall, our results indicate that XIST delivered by monocyte-derived Exos aggravates ALI via regulating the miR-448-5p/HMGB2 signaling axis.
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Lesão Pulmonar Aguda , MicroRNAs , RNA Longo não Codificante , Camundongos , Animais , MicroRNAs/genética , MicroRNAs/metabolismo , Proteína HMGB2/genética , Monócitos/metabolismo , Lipopolissacarídeos/efeitos adversos , Fatores de Transcrição , Lesão Pulmonar Aguda/induzido quimicamente , Lesão Pulmonar Aguda/genética , Lesão Pulmonar Aguda/terapia , RNA Longo não Codificante/genéticaRESUMO
Nonalcoholic fatty liver disease (NAFLD) is related to the production of reactive oxygen species (ROS) and oxidative stress, so antioxidant treatment can prevent its further development. Ultrasmall copper-based nanoparticles (CuNPs) have shown multiple enzyme-like activities for scavenging oxygen species, providing a new strategy for the treatment of inflammatory diseases. Resveratrol (Res), a natural polyphenol compound, has attracted much attention due to its ability to inhibit oxidative stress. We therefore aimed to first combine these two agents for the treatment of NAFLD. However, due to the poor water solubility and stability of Res, which is easily metabolized in the intestine, the development of a stable and effective carrier became the key to achieving a synergistic effect. Liver-targeted nanocarriers loaded with bioactive compounds may provide a more effective approach for the treatment of NAFLD. Therefore, we developed a novel ultrasonic nanobubble carrying nucleic acid aptamers with liver targeting properties, which has the advantages of a small molecular weight, no immunogenicity, a low cost of synthesis, and high stability through chemical modification. Res and the ultrasmall CuNPs were specifically delivered to liver tissue to maximize therapeutic efficiency. This study found that the combination of these two components can effectively treat inflammation in NAFLD and suggested that liver-targeted NAFLD-specific aptamer-mediated targeted ultrasound nanobubbles that can simultaneously deliver Res and CuNPs may be a safe and effective new platform for NAFLD and other liver diseases.
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The representative morphological features of pyroptosis are excessive cell swelling and subsequent membrane rupture. However, the mechanism underlying the cell's inherent inability to regulate volume during the progression of pyroptosis is poorly understood. In the current study, we found that both volume-activated chloride currents (Icl, vol) and the regulatory volume decrease (RVD) were markedly decreased in bone marrow-derived macrophages (BMDMs) undergoing pyroptosis induced by lipopolysaccharides (LPS) and nigericin. The inhibition of ICl, vol and RVD by the chloride channel blockers, tamoxifen or DCPIB, led to the emergence of pyroptosis-like phenotypes such as activated-caspase-1, pyroptotic-body-like bubbles, and a fried-egg-like appearance. Moreover, the expression of the volume-activated chloride channel (VRAC) constituent protein Leucine-Rich Repeat-Containing 8A (LRRC8A) was significantly down-regulated in pyroptotic BMDMs treated with LPS and nigericin. The silencing of LRRC8A expression by small interfering RNA (si)-LRRC8A transfection not only reduced ICl, vol and RVD, but also caused BMDMs to show pyroptosis-like manifestations such as activated-caspase-1, membrane bubbles, and have a fried-egg-like appearance. These results reveal a new mechanism for the loss of volume regulation in the process of pyroptotic cell swelling and strongly suggest that a functional deficiency of VRAC/LRRC8A plays a key role in this disorder.
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Canais de Cloreto/metabolismo , Lipopolissacarídeos/toxicidade , Nigericina/toxicidade , Piroptose/efeitos dos fármacos , Animais , Antibacterianos/toxicidade , Biomarcadores , Ciclopentanos/farmacologia , Antagonistas de Estrogênios/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Indanos/farmacologia , Macrófagos , Masculino , Proteínas de Membrana , Camundongos , Camundongos Endogâmicos C57BL , RNA Interferente Pequeno , Tamoxifeno/farmacologiaRESUMO
Nanobubbles (NBs) are considered to be a new generation of ultrasound-responsive nanocarriers that can effectively target tumors, accurately release multi-drugs at desired locations, as well as simultaneously perform diagnosis and treatment. In this study, we designed theranostic NBs (FTY720@SPION/PFP/RGD-NBs) composed of RGD-modified liposomes as the shell, and perflenapent (PFP), superparamagnetic iron oxide nanoparticles (SPION), and fingolimod (2-amino-2[2-(4-octylphenyl)ethyl]-1,3-propanediol, FTY720) encapsulated as the core. The prepared FTY720@SPION/PFP/RGD-NBs were black spheres with a diameter range of 160-220 nm, eligible for enhanced permeability and retention (EPR) effects. The calculated average drug loading efficiency (LE) and encapsulation efficiency (EE) of the FTY720@SPION/PFP/RGD-NBs were 9.18 ± 0.61% and 88.26 ± 2.31%, respectively. With the promotion of low-intensity focused ultrasound (LIFU), the amount and the rate of FTY720 released from the prepared NB complex were enhanced when compared to the samples without LIFU treatment. In vitro magnetic resonance imaging (MRI) trials showed that the prepared FTY720@SPION/PFP/RGD-NBs had a high relaxation rate and MRI T2-weighted imaging (T2WI) scanning sensitivity conditions. The cell viability studies demonstrated that both HepG2 and Huh7 cells co-cultured with FTY720@SPION/PFP/RGD-NB (100 µg mL-1) + LIFU treatment had the lowest survival rate compared with the other groups at 24 h and 48 h, showing that FTY720@SPION/PFP/RGD-NB had the strongest anti-tumor efficiency among the prepared NBs. The cytotoxicity study also demonstrated that the prepared NBs had low toxicity to normal fibroblast 3T3 cells. Cellular uptake studies further indicated that both LIFU treatment and RGD modification could effectively improve the tumor-targeted effects, thereby enhancing the antitumor efficacy. The qRT-PCR results indicated that LIFU-mediated FTY720@SPION/PFP/RGD-NB could significantly cause the activation of Caspase3, Caspase9 and p53 compared to the control group, inducing HepG2 apoptosis. These results together indicated that FTY720@SPION/PFP/RGD-NBs combined with LIFU may serve as a multifunctional drug delivery platform for hepatocellular carcinoma treatment and provide a new strategy for tumor visualization by MRI.
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The present study aimed to investigate the role of caprin-1 in liver cancer and its association with the clinicopathological features and prognosis of liver cancer, as well as the underlying mechanism of caprin-1 function. Caprin-1 expression levels in a tissue microarray containing 40 liver cancer tissues, 10 peritumoral tissues and 20 normal liver tissues were analyzed using immunohistochemistry. The clinical data of 154 patients with liver cancer were also collected from The Cancer Genome Atlas database. Kaplan-Meier analysis and a Cox proportional hazards regression model were used to assess the association between caprin-1 expression levels and survival in patients with liver cancer. The effects of caprin-1 knockdown on the mRNA levels of cyclin D1 and cyclin D2 as well as the proliferation, invasion and migration of HepG2 cells were also investigated. The expression level of caprin-1 in liver cancer tissues was significantly higher compared with normal liver tissues or cells (P<0.01). High caprin-1 expression levels were associated with advanced clinical stage (P<0.001) and enhanced tumor invasion (P<0.001). Kaplan-Meier analysis showed that the overall survival time and disease-free survival time in patients with liver cancer with high caprin-1 expression were significantly shorter compared with patients with low caprin-1 expression levels (P=0.002 and P=0.033, respectively). The Cox proportional hazards regression model showed that high caprin-1 expression levels were an independent prognostic factor for liver cancer (P<0.001). Knockdown of caprin-1 in HepG2 cells significantly downregulated mRNA expression levels of cyclin D1 and cyclin D2, inhibited cell proliferation and invasion and the cells were arrested at G0/G1 phase. In conclusion, caprin-1 may be a novel prognostic indicator for patients with liver cancer.
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Platelet dysfunction plays a critical role in vascular complications of type 2 diabetes mellitus (T2DM). But the relationship between platelet hyperactivity and its energy metabolic process remains unclear. This study was designed to explore alterations of platelet mitochondrial ATP production and the possible mechanism. A total of 39 T2DM patients without macrovascular and microvascular complications and 32 normal controls were fasting sampled. Platelet ATP content was measured by a high performance liquid chromatograph (HPLC). The flow cytometry technique was adopted to evaluate mitochondrial membrane potential (DeltaPsim), the stored force for platelet ATP production. Consequently, T2DM patients exhibited obvious hyperglycemia, hyperlipidemia and hypertension, but normal platelet morphology. Platelet ATP content was significantly higher in T2DM (0.032 +/- 0.010 micromol/10(9) platelets versus 0.017 +/- 0.006 micromol/10(9) platelets, p < 0.001) than in the control group. Interestingly, DeltaPsim was markedly decreased in T2DM patients (0.79 +/- 0.18 versus 2.70 +/- 1.03, p < 0.001) compared with normal controls. For whole subjects, a stepwise regression showed that plasma glycated hemoglobin A1c (HbA1c) level positively correlated to platelet ATP content (beta = 0.552, 95% CI = 0.072-1.451), and fasting plasma glucose (FPG) level was negatively correlated to DeltaPsim (beta = -0.372, 95% CI = -0.471 to -0.089). These data support that hyperglycemia of T2DM promotes platelet mitochondria to generate more ATP, but decreases platelet mitochondrial potential. The discordance between them requires further researches to elucidate.
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Trifosfato de Adenosina/metabolismo , Plaquetas/metabolismo , Diabetes Mellitus Tipo 2/sangue , Potencial da Membrana Mitocondrial/fisiologia , Idoso , Animais , Plaquetas/citologia , Feminino , Humanos , Hiperglicemia/metabolismo , Masculino , Pessoa de Meia-IdadeRESUMO
Based on the results of phenotypic features, phylogenetic similarity of 16S rRNA gene sequences and BIOLOG test, a soil bacterium was identified as Bacillus sp. DM-1. Using either growing cells or a cell-free extract, it transformed parathion and methyl parathion to amino derivatives by reducing the nitro group. Pesticide transformation by a cell-free extract was specifically inhibited by three nitroreductase inhibitors, indicating the presence of nitroreductase activity. The nitroreductase activity was NAD(P)H-dependent, O(2)-insensitive, and exhibited the substrate specificity for parathion and methyl parathion. Reductive transformation significantly decreased the toxicity of pesticides.
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Bacillus/metabolismo , Metil Paration/metabolismo , Paration/metabolismo , Biodegradação Ambiental , OxirreduçãoRESUMO
OBJECTIVE: To investigate the effect of Rehmannia glutinosa water extraction (RGL) on adipose metabolic disorder and gene expression of resistin in type 2 diabetes mellitus rats. METHOD: The wistar rats model of 2-DM were induced by high calorie feeding and small dose injection of STZ. Rats were randomly divided into diabetic model group, diabetic model treated with RGL (2.4 g x kg(-1) x d(-1)), RGL (1.2 g x kg(-1) x d(-1)), RGL (0. 6 g x kg(-1) x d(-1)) and normal control group. The levels of FPG, FINS, TG, HDL, LDL, CH and IR were measured, and the mRNA expression of resistin was determined by RT-PCR, the protein expression measured by SDS-PAGD at the end of 8 weeks. RESULT: The gene expression of resistin in RGL group were lower than that of diabetic model (P < 0.01). The levels of FPG, FINS, TG, LDL, CH, IR in RGL group were lower than that of diabetic model (P < 0.05), and HDL were higher (P < 0.05). CONCLUDE: RGL can improve insulin resistance in the experimental 2-DM rats, can effectively ameliorate adipose metabolic disturbance and decline IR and FINS by increasing the gene expression of resistin.
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Diabetes Mellitus Tipo 2/patologia , Medicamentos de Ervas Chinesas/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Resistência à Insulina , Rehmannia/química , Resistina/genética , Animais , Glicemia/metabolismo , Colesterol/sangue , Diabetes Mellitus Experimental/sangue , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Experimental/patologia , Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/genética , Medicamentos de Ervas Chinesas/isolamento & purificação , Eletroforese em Gel de Poliacrilamida , Feminino , Hipoglicemiantes/isolamento & purificação , Hipoglicemiantes/farmacologia , Insulina/sangue , Masculino , Plantas Medicinais/química , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Distribuição Aleatória , Ratos , Ratos Wistar , Resistina/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Triglicerídeos/sangueRESUMO
Gene therapy, particularly microRNA (miRNA), is a promising candidate in the treatment of cancer; however, it is challenging to develop gene delivery systems. Ultrasound microbubbles have been used for gene delivery with excellent results. The present study aimed to investigate the transfection efficiency of HepG2 cells using ultrasound microbubbles. The effects of three miRNAs (miR-21, miR-221 and miR-199a) on HepG2 cells were also determined by performing ultrasound microbubble-mediated gene transfection. Three recombinant plasmids containing anti-miR-21, anti-miR-221 and miR-199a were fused with enhanced green fluorescent protein. For the transfection of genes, the type of contrast agent, the concentration of microbubble contrast agent and the exposure intensity of ultrasound were optimized. The expression of miRNAs was detected using reverse transcription-polymerase chain reaction. To determine the effect of anti-miR-21, anti-miR-221 and miR-199a on HepG2 cells, MTT, cell cycle analysis and Annexin V-PE/7-ADD apoptosis assays were performed. The optimal condition was 10% sulfur hexafluoride microbubbles at an ultrasound frequency of 2.0 MHz and mechanical index of 0.28. When cells were transfected with three recombinant plasmids using ultrasound microbubbles, there was significant downregulation of miR-21 and miR-221 and upregulation of miR-199a (P<0.05). All three treatments inhibited cell proliferation and promoted the apoptosis of cells. The present data indicated that the delivery of anti-miR-21, anti-miR-221 and miR-199a may be mediated by ultrasound microbubble contrast agents. With this approach, cell proliferation may be effectively inhibited and cell apoptosis may be induced. These are novel cancer therapy targets.
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An effective chlorpyrifos-degrading bacterium (named strain YC-1) was isolated from the sludge of the wastewater treating system of an organophosphorus pesticides manufacturer. Based on the results of phenotypic features, phylogenetic similarity of 16S rRNA gene sequences and BIOLOG test, strain YC-1 was identified as the genus Stenotrophomonas. The isolate utilized chlorpyrifos as the sole source of carbon and phosphorus for its growth and hydrolyzed chlorpyrifos to 3,5,6-trichloro-2-pyridinol. Parathion, methyl parathion, and fenitrothion also could be degraded by strain YC-1 when provided as the sole source of carbon and phosphorus. The gene encoding the organophosphorus hydrolase was cloned using a PCR cloning strategy based on the known methyl parathion degrading (mpd) gene of Plesiomonas sp. M6. Sequence blast result indicated this gene has 99% similar to mpd. The inoculation of strain YC-1 (10(6) cells g(-1)) to soil treated with 100 mg kg(-1) chlorpyrifos resulted in a higher degradation rate than in noninoculated soils. Theses results highlight the potential of this bacterium to be used in the cleanup of contaminated pesticide waste in the environment.