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Myriad physiological and pathogenic processes are governed by protein levels and modifications. Controlled protein activity perturbation is essential to studying protein function in cells and animals. Based on Trim-Away technology, we screened for truncation variants of E3 ubiquitinase Trim21 with elevated efficiency (ΔTrim21) and developed multiple ΔTrim21-based targeted protein-degradation systems (ΔTrim-TPD) that can be transfected into host cells. Three ΔTrim-TPD variants are developed to enable chemical and light-triggered programmable activation of TPD in cells and animals. Specifically, we used ΔTrim-TPD for (1) red-light-triggered inhibition of HSV-1 virus proliferation by degrading the packaging protein gD, (2) for chemical-triggered control of the activity of Cas9/dCas9 protein for gene editing, and (3) for blue-light-triggered degradation of two tumor-associated proteins for spatiotemporal inhibition of melanoma tumor growth in mice. Our study demonstrates that multiple ΔTrim21-based controllable TPD systems provide powerful tools for basic biology research and highlight their potential biomedical applications.
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Sistemas CRISPR-Cas , Edición Génica , Ratones , Animales , Proteína 9 Asociada a CRISPR/genética , Proteína 9 Asociada a CRISPR/metabolismo , Proteínas/metabolismo , Proteolisis , Mamíferos/metabolismoRESUMEN
Chimeric antigen receptor (CAR)-engineered T cell therapies have been recognized as powerful strategies in cancer immunotherapy; however, the clinical application of CAR-T is currently constrained by severe adverse effects in patients, caused by excessive cytotoxic activity and poor T cell control. Herein, we harnessed a dietary molecule resveratrol (RES)-responsive transactivator and a transrepressor to develop a repressible transgene expression (RESrep) device and an inducible transgene expression (RESind) device, respectively. After optimization, these tools enabled the control of CAR expression and CAR-mediated antitumor function in engineered human cells. We demonstrated that a resveratrol-repressible CAR expression (RESrep-CAR) device can effectively inhibit T cell activation upon resveratrol administration in primary T cells and a xenograft tumor mouse model. Additionally, we exhibit how a resveratrol-inducible CAR expression (RESind-CAR) device can achieve fine-tuned and reversible control over T cell activation via a resveratrol-titratable mechanism. Furthermore, our results revealed that the presence of RES can activate RESind-CAR T cells with strong anticancer cytotoxicity against cells in vitro and in vivo. Our study demonstrates the utility of RESrep and RESind devices as effective tools for transgene expression and illustrates the potential of RESrep-CAR and RESind-CAR devices to enhance patient safety in precision cancer immunotherapies.
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Citotoxicidad Inmunológica/inmunología , Inmunoterapia Adoptiva/métodos , Leucemia Eritroblástica Aguda/inmunología , Activación de Linfocitos/inmunología , Receptores de Antígenos de Linfocitos T/inmunología , Receptores Quiméricos de Antígenos/inmunología , Linfocitos T/inmunología , Animales , Apoptosis , Proliferación Celular , Modelos Animales de Enfermedad , Femenino , Humanos , Leucemia Eritroblástica Aguda/genética , Leucemia Eritroblástica Aguda/metabolismo , Leucemia Eritroblástica Aguda/terapia , Ratones , Ratones Endogámicos NOD , Ratones SCID , Células Tumorales Cultivadas , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Diabetes affects almost half a billion people, and all individuals with type 1 diabetes (T1D) and a large portion of individuals with type 2 diabetes rely on self-administration of the peptide hormone insulin to achieve glucose control. However, this treatment modality has cumbersome storage and equipment requirements and is susceptible to fatal user error. Here, reasoning that a cell-based therapy could be coupled to an external induction circuit for blood glucose control, as a proof of concept we developed far-red light (FRL)-activated human islet-like designer (FAID) cells and demonstrated how FAID cell implants achieved safe and sustained glucose control in diabetic model mice. Specifically, by introducing a FRL-triggered optogenetic device into human mesenchymal stem cells (hMSCs), which we encapsulated in poly-(l-lysine)-alginate and implanted subcutaneously under the dorsum of T1D model mice, we achieved FRL illumination-inducible secretion of insulin that yielded improvements in glucose tolerance and sustained blood glucose control over traditional insulin glargine treatment. Moreover, the FAID cell implants attenuated both oxidative stress and development of multiple diabetes-related complications in kidneys. This optogenetics-controlled "living cell factory" platform could be harnessed to develop multiple synthetic designer therapeutic cells to achieve long-term yet precisely controllable drug delivery.
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Diabetes Mellitus Tipo 1 , Diabetes Mellitus Tipo 2 , Animales , Glucemia , Diabetes Mellitus Tipo 1/terapia , Humanos , Insulina/metabolismo , Secreción de Insulina , RatonesRESUMEN
Di-(2-ethylhexyl) phthalate (DEHP) and mono-2-ethylhexyl phthalate (MEHP) can induce hepatic lipid metabolism disorders, while the molecular mechanism still remain unknown. We aim to explore the underlying mechanism of Notch signaling pathway on hepatic lipid accumulation induced by DEHP/MEHP. A total of 40 male wistar rats were exposed to DEHP (0, 5, 50, and 500 mg/kg/d) for 8 weeks, BRL-3A hepatocytes were exposed to MEHP (0, 10, 50, 100, and 200 µM) for 24 h. About 50 µM DAPT and 100 µg/mL Aspirin were used to inhibit Notch pathway and prevent inflammation, respectively. Real-Time PCR was performed to detect the mRNA expression, western blot and immunofluorescence were used to detect the protein expression. Lipids and inflammatory factors levels were determined by commercial kits. The results showed that DEHP/MEHP promoted the expression of Notch pathway molecules and lipids accumulation in rat livers/BRL-3A cells. The up-regulated Notch receptors were correlated with the TG levels in the rat liver. MEHP increased the levels of IL-8 and IL-1ß. The lipids levels were reduced after anti-inflammation. The inhibition of Notch pathway reversed the elevation of inflammation and lipid accumulation caused by MEHP. In conclusion, this study demonstrated that DEHP/MEHP led to lipid accumulation in hepatocytes by up-regulating Notch pathway and the inflammation might play a key role in the process.
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Dietilhexil Ftalato , Ratas , Animales , Masculino , Dietilhexil Ftalato/metabolismo , Hígado/metabolismo , Ratas Wistar , Transducción de Señal , Inflamación , LípidosRESUMEN
Colorectal polyp has been considered as the precancerous lesion of colorectal cancer, to which serum lipid levels are closely related. At present, there is no consensus on the relationship between colorectal polyps and serum lipid levels. We performed a meta-analysis to explore the effects of lipid levels on colorectal polyps. Relevant articles published from 2000 to 2020 were searched in PubMed, Web of Science, EMBASE, and Cochrane Library databases. The mean value and SD of serum lipid indexes and body mass index in colorectal polyps groups and control groups were extracted from the included articles. Combined weighted mean differences (WMDs) and 95% confidence intervals (CIs) were calculated to assess the effect size of serum lipid levels on colorectal polyps. The publication bias of the included studies were assessed based on the Egger test. Thirty-seven articles containing 19,464 cases and 63,979 controls were included. There were no significant publication bias. The levels of high-density lipoprotein cholesterol in the cases were lower than those in the controls (WMD: -2.589 mg/dL, 95% CI: -3.273, -1.906). While the levels of triglyceride (WMD: 16.933 mg/dL, 95% CI: 13.131, 20.736), total cholesterol (WMD: 5.561 mg/dL, 95% CI: 3.477, 7.645), low-density lipoprotein cholesterol (WMD: 3.109 mg/dL, 95% CI: 0.859, 5.359) and body mass index (WMD: 0.747 mg/dL, 95% CI: 0.588, 0.906) were higher in the cases. Colorectal polyps were associated with serum lipid levels and obesity. Hyperlipidemia and obesity may be the risk factors for colorectal polyps.
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Pólipos del Colon , HDL-Colesterol , LDL-Colesterol , Humanos , Obesidad/complicaciones , TriglicéridosRESUMEN
Atmospheric PM2.5 can induce airway inflammation and mucin secretion. MUC5B is required for airway defense. However, the research on the role of MUC5B in airway inflammation induced by atmospheric PM2.5 remains limited. This study was designed to explore the role of MUC5B in airway inflammation induced by atmospheric PM2.5. In vivo, Wistar rats were exposed to 0, 1.5, 7.5, 37.5 mg/ kg PM2.5 saline suspension via intratracheal instillation. HE staining and AB-PAS staining were used to observe the airway inflammation and goblet cell hyperplasia. In vitro, normal A549 cells and MUC5B-knockdown A549 cells were exposed to 0, 100, 200 and 400 µg/mL PM2.5 for 6 h, 12 h, 24 h and 48 h. ELISA was used to measure the levels of TNF-α and IL-1ß in serum and bronchoalveolar lavage fluid of rats and in cell culture. Real time-PCR and ELISA were used to quantify the mRNA and protein levels of MUC5B in trachea and lung of rats and in A549 cells. PM2.5 could cause the infiltration of inflammatory cells and increase the mucus secretions and goblet cell metaplasia. MUC5B is related to rats' airway inflammation induced by PM2.5. A549 cells exposed to PM2.5 in higher concentration and longer time, the protein level of MUC5B was significantly increased, while the levels of IL-1ß, TNF-α and MUC5B mRNA were significantly decreased. Compared with normal A549 cells, the levels of IL-1ß and TNF-α were significantly higher in Muc5b-knockdown cells. Atmospheric PM2.5 can induce airway inflammation and mucin secretion. MUC5B played a critical role in controlling the inflammatory response induced by PM2.5.
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Inflamación/metabolismo , Mucina 5B/metabolismo , Material Particulado/toxicidad , Células A549 , Animales , Líquido del Lavado Bronquioalveolar/química , Femenino , Humanos , Interleucina-1beta/sangre , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Pulmón/metabolismo , Masculino , Mucina 5B/genética , Ratas , Ratas Wistar , Factor de Necrosis Tumoral alfa/sangre , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Hepatogenous diabetes is a complex disease that is typified by the simultaneous presence of type 2 diabetes and many forms of liver disease. The chief pathogenic determinant in this pathophysiological network is insulin resistance (IR), an asymptomatic disease state in which impaired insulin signaling in target tissues initiates a variety of organ dysfunctions. However, pharmacotherapies targeting IR remain limited and are generally inapplicable for liver disease patients. Oleanolic acid (OA) is a plant-derived triterpenoid that is frequently used in Chinese medicine as a safe but slow-acting treatment in many liver disorders. Here, we utilized the congruent pharmacological activities of OA and glucagon-like-peptide 1 (GLP-1) in relieving IR and improving liver and pancreas functions and used a synthetic-biology-inspired design principle to engineer a therapeutic gene circuit that enables a concerted action of both drugs. In particular, OA-triggered short human GLP-1 (shGLP-1) expression in hepatogenous diabetic mice rapidly and simultaneously attenuated many disease-specific metabolic failures, whereas OA or shGLP-1 monotherapy failed to achieve corresponding therapeutic effects. Collectively, this work shows that rationally engineered synthetic gene circuits are capable of treating multifactorial diseases in a synergistic manner by multiplexing the targeting efficacies of single therapeutics.
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Diabetes Mellitus Tipo 2/etiología , Diabetes Mellitus Tipo 2/terapia , Hepatopatías/complicaciones , Animales , Ingeniería Celular , Línea Celular , Diabetes Mellitus Tipo 2/diagnóstico , Diabetes Mellitus Tipo 2/metabolismo , Modelos Animales de Enfermedad , Diseño de Fármacos , Regulación de la Expresión Génica/efectos de los fármacos , Redes Reguladoras de Genes/efectos de los fármacos , Ingeniería Genética , Péptido 1 Similar al Glucagón/farmacología , Péptido 1 Similar al Glucagón/uso terapéutico , Humanos , Resistencia a la Insulina , Hepatopatías/metabolismo , Masculino , Ratones , Ratones Transgénicos , Ácido Oleanólico/farmacología , Ácido Oleanólico/uso terapéutico , Biología SintéticaRESUMEN
In order to explore the feasibility of applying magnetic detection electrical impedance tomography(MDEIT) on respiratory monitoring, aiming at the forward problem of magnetic detection electrical impedance tomography, we calculated the electric potential and current density distribution inside the imaging object by using the finite element method. We then got magnetic induction intensity outside the object at the end of exhaling and inhaling according to Biot-Savart's law. The results showed that the magnetic induction intensity at the end of inhaling was 8.875%,less than that at the end of exhaling. By the simulation results, we could understand the difference of magnetic induction intensity value surrounding the lung at the end of exhaling and inhaling due to the change of lung volume and electrical conductivity distribution better. Our research laid the foundation for the late image reconstruction and clinical disease detection.
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Magnetismo , Algoritmos , Simulación por Computador , Impedancia Eléctrica , Humanos , Interpretación de Imagen Asistida por Computador/métodos , Procesamiento de Imagen Asistido por Computador/métodos , Fantasmas de Imagen , Tomografía/métodosRESUMEN
Despite the advantages of using lightweight and non-corrosive carbon fiber reinforced polymer (CFRP) reinforcements in concrete structures, their widespread adoption has been limited due to concerns regarding the brittle failure of CFRP rupture and its relatively softer load-deflection stiffness. This work offers logical solutions to these two crucial problems: using aggregate coating to strengthen the CFRP-concrete bond and ultimately the load-deflection stiffness, and using CFRP-concrete debonding propagation to create pseudo-ductile behavior. Subsequently, the concrete cracking behavior, the apparent CFRP modulus with aggregates, and the post-peak capacity and deflection of three-dimensional (3D) CFRP-reinforced concrete are all described by equations derived from experiments. These formulas will be helpful in the future design of non-prismatic concrete components for low-impact building projects. The potential of this innovative design scheme in terms of increased capacity and deflections with less concrete material is demonstrated through comparisons between non-prismatic CFRP-reinforced concrete and measured steel reinforced equivalency.
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Di-(2-ethylhexyl) phthalate (DEHP) is one of the most widely used plasticizers. Many studies focus on the impact of continuous exposure to DEHP on humans and ecosystems. In this study, the bibliometric analysis of DEHP and its metabolites research was conducted to assess the research performances, hotspot issues, and trends in this field. The data was retrieved from a Web of Science Core Collection online database. VOSviewer 1.6.18 was used to analyze. A total of 4672 publications were collected from 1975 to 2022 October 21. The number of publications and citations increased annually in the last decades. China had the largest number of publications, and the USA had the highest co-authorship score. The most productive and most frequently cited institutions were the Chinese Academy of Sciences and the Centers for Disease Control & Prevention (USA), respectively. The journal with the most publications was the Science of Total Environment, and the most cited one was the Environmental Health Perspectives. The most productive and cited author was Calafat A. M. (USA). The most cited reference was "Phthalates: toxicology and exposure." Four hotspot issues were as follows: influences of DEHP on the organisms and its possible mechanisms, assessment of DEHP exposure to the human and its metabolism, dynamics of DEHP in external environments, and indoor exposure of DEHP and health outcomes. The research trends were DNOP, preterm birth, gut microbiota, microplastics, lycopene, hypertension, and thyroid hormones. This study can provide researchers with new ideas and decision-makers with reference basis to formulate relevant policies.
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Bibliometría , Dietilhexil Ftalato , Humanos , Plastificantes , Ácidos Ftálicos , ChinaRESUMEN
Synthetic biology applications require finely tuned gene expression, often mediated by synthetic transcription factors (sTFs) compatible with the human genome and transcriptional regulation mechanisms. While various DNA-binding and activation domains have been developed for different applications, advanced artificially controllable sTFs with improved regulatory capabilities are required for increasingly sophisticated applications. Here, in mammalian cells and mice, we validate the transactivator function and homo-/heterodimerization activity of the plant-derived phytochrome chaperone proteins, FHY1 and FHL. Our results demonstrate that FHY1/FHL form a photosensing transcriptional regulation complex (PTRC) through interaction with the phytochrome, ΔPhyA, that can toggle between active and inactive states through exposure to red or far-red light, respectively. Exploiting this capability, we develop a light-switchable platform that allows for orthogonal, modular, and tunable control of gene transcription, and incorporate it into a PTRC-controlled CRISPRa system (PTRCdcas) to modulate endogenous gene expression. We then integrate the PTRC with small molecule- or blue light-inducible regulatory modules to construct a variety of highly tunable systems that allow rapid and reversible control of transcriptional regulation in vitro and in vivo. Validation and deployment of these plant-derived phytochrome chaperone proteins in a PTRC platform have produced a versatile, powerful tool for advanced research and biomedical engineering applications.
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Luz , Chaperonas Moleculares , Fitocromo , Animales , Humanos , Ratones , Fitocromo/metabolismo , Fitocromo/genética , Chaperonas Moleculares/metabolismo , Chaperonas Moleculares/genética , Regulación de la Expresión Génica/efectos de la radiación , Transcripción Genética/efectos de la radiación , Células HEK293 , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Factores de Transcripción/metabolismo , Factores de Transcripción/genética , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genéticaRESUMEN
Gene therapies provide treatment options for many diseases, but the safe and long-term control of therapeutic transgene expression remains a primary issue for clinical applications. Here, we develop a muscone-induced transgene system packaged into adeno-associated virus (AAV) vectors (AAVMUSE) based on a G protein-coupled murine olfactory receptor (MOR215-1) and a synthetic cAMP-responsive promoter (PCRE). Upon exposure to the trigger, muscone binds to MOR215-1 and activates the cAMP signaling pathway to initiate transgene expression. AAVMUSE enables remote, muscone dose- and exposure-time-dependent control of luciferase expression in the livers or lungs of mice for at least 20 weeks. Moreover, we apply this AAVMUSE to treat two chronic inflammatory diseases: nonalcoholic fatty liver disease (NAFLD) and allergic asthma, showing that inhalation of muscone-after only one injection of AAVMUSE-can achieve long-term controllable expression of therapeutic proteins (ΔhFGF21 or ΔmIL-4). Our odorant-molecule-controlled system can advance gene-based precision therapies for human diseases.
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Alprostadil , Cicloparafinas , Ratones , Humanos , Animales , Alprostadil/metabolismo , Transgenes , Cicloparafinas/metabolismo , Odorantes , Receptores Acoplados a Proteínas G/metabolismo , Dependovirus/genética , Vectores GenéticosRESUMEN
Plastics in the environment can be degraded and even broken into pieces under the action of natural factors, and the degraded products with a particle size of less than 5 mm are called microplastics (MPs). MPs exist in a variety of environmental media that come into contact with the human body. It can enter the body through environmental media and food chains. At present, there are many studies investigating the damage of MPs to marine organisms and mammals. The liver is the largest metabolizing organ and plays an important role in the metabolism of MPs in the body. However, there is no available systematic review on the toxic effects of MPs on the liver. This paper summarizes the adverse effects and mechanisms of MPs on the liver, by searching the literature and highlighting the studies that have been published to date, and provides a scenario for the liver toxicity caused by MPs.
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Background: The benefit of cold exposure for humans against obesity has brought the energy metabolism and activity of brown adipose tissue (BAT) induced by cold into focus. But the results are inconsistent. This review is aimed to systematically explore the effect of cold exposure on the activity of BAT and energy metabolism in humans. Methods: We searched relevant papers that were published from 1990 to 2021 and were cited in PubMed Central, Web of science, Embase and Cochrane Library databases to conduct this systematic review and meta-analysis. Energy metabolism, BAT volume, BAT activity and non-esterified fatty acids (NEFA) data reported in eligible researches were extracted. Meta-analysis was applied to combine the mean difference or standard mean difference with their 95% confidence intervals (95%CI). RevMan 5.3 software was used for meta-analysis and evaluating the risk of bias. Stata 16.0 was used for evaluating the publication bias. Results: Ten randomized controlled trials were included in meta-analysis. Compared with human exposed in room temperature at 24°C, the energy expenditure (EE) was increased after acute cold exposure at 16â¼19°C (Z = 7.58, p < 0.05, mean different = 188.43kal/d, 95% CI = 139.73-237.13); BAT volume (Z = 2.62, p < 0.05; standard mean different = 0.41, 95% CI = 0.10-0.73); BAT activity (Z = 2.05, p = 0.04, standard mean difference = 1.61, 95% CI = 0.07-3.14) and the intake of BAT NEFA (Z = 2.85, p < 0.05; standard mean different = 0.53, 95% CI = 0.17-0.90) also increased. Conclusion: Acute cold exposure could improve the energy expenditure and BAT activity in adults, which is beneficial for human against obesity.
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In recent years, the incidence of lipid metabolism disorders in adolescents has gradually increased, and the effects of DEHP on lipid metabolism have received widespread attention. In this study, 463 adolescents aged 16-19 years were enrolled as subjects. This study analyzed the associations between the urinary levels of DEHP metabolites (MEHP, MEOHP, MEHHP, MECPP, MCMHP, and ∑DEHP) and BMI, WHR, WtHR, VAI, LAP, the plasma levels of lipids (TC, TG, HDL-C, and LDL-C), and the peripheral blood leukocyte mRNA levels of SREBP-2, SR-BI, LDLR, and NR1H3. Animal experiments were performed to confirm and expand findings. Wistar rats were administered DEHP at 0, 5, 50, and 500 mg/kg/d for 8 weeks. The serum and liver levels of TC, TG, HDL-C, and LDL-C, and the liver mRNA and protein levels of SREBP-2, SR-BI, LDLR, and NR1H3 were measured. The results showed that WHR, VAI, and LAP were significantly positively associated with the urinary levels of MECPP and ∑DEHP; the plasma HDL-C level was significantly negatively associated with the levels of MECPP, MCMHP and ∑DEHP; the peripheral blood leukocyte mRNA levels of SREBP-2, NR1H3, and LDLR were significantly positively correlated with the MCMHP level; and the SR-BI mRNA level was significantly positively correlated with the levels of MECPP and MCMHP in adolescents. Moreover, the results of animal experiments showed that DEHP exposure significantly increased the serum levels of TC, HDL-C, and LDL-C in 500 mg/kg/d group, as well as the liver levels of TC and HDL-C, up-regulated SREBP-2 mRNA and protein expression in 50 and 500 mg/kg/d groups. DEHP exposure significantly down-regulated SR-BI and NR1H3 protein expression in the liver of the 500 mg/kg/d group rats. Our findings indicate that DEHP exposure can affect lipid metabolism in adolescents by regulating the expression of lipid metabolism-related genes.
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Dietilhexil Ftalato , Ácidos Ftálicos , Adolescente , Animales , Dietilhexil Ftalato/toxicidad , Exposición a Riesgos Ambientales , Humanos , Metabolismo de los Lípidos , Ratas , Ratas WistarRESUMEN
The CRISPR-Cas12a has been harnessed as a powerful tool for manipulating targeted gene expression. The possibility to manipulate the activity of CRISPR-Cas12a with a more precise spatiotemporal resolution and deep tissue permeability will enable targeted genome engineering and deepen our understanding of the gene functions underlying complex cellular behaviors. However, currently available inducible CRISPR-Cas12a systems are limited by diffusion, cytotoxicity, and poor tissue permeability. Here, we developed a far-red light (FRL)inducible CRISPR-Cas12a (FICA) system that can robustly induce gene editing in mammalian cells, and an FRL-inducible CRISPR-dCas12a (FIdCA) system based on the protein-tagging system SUperNova (SunTag) that can be used for gene activation under light-emitting diodebased FRL. Moreover, we show that the FIdCA system can be deployed to activate target genes in mouse livers. These results demonstrate that these systems developed here provide robust and efficient platforms for programmable genome manipulation in a noninvasive and spatiotemporal fashion.
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Fine particulate matter (PM2.5) exposure can cause the injury of vascular endothelial cells by inflammatory response. CD40 works in inflammation of endothelial cells and it may be regulated by the miRNAs. This study aimed to clarify the role and mechanism of CD40 and miR-145-5p in PM2.5-induced injury of human umbilical vein endothelial cells (HUVECs). HUVECs were treated with different concentrations of PM2.5 exposure (0, 100, 200, 400 µg/mL) for 24 h. The si-RNA was used for CD40 gene silencing (0, 200 µg/mL PM2.5, siRNA-CD40 and siRNA-CD40 + 200 µg/mL PM2.5). Mimics was used for overexpression of miR-145-5p (0, 200 µg/mL PM2.5, mimics and mimics+200 µg/mL PM2.5). The cell viability of HUVECs was detected with Cell Counting Kit8 (CCK8) kit. The level of cell apoptosis was detected by flow cytometry. The inflammation-related factor including interleukin-1ß (IL-1ß), interleukin-18 (IL-18), tumor necrosis factor α (TNF-α) and C1q complement/tumor necrosis factor (TNF)-associated proteins9 (CTRP9) were tested with enzyme-linked immunosorbent assay (ELISA) kits. The mRNA and protein expression levels of CD40, CD40L, caspase1, NLRP3 (Nod-like receptor family pyrin domain-containing 3) and IKKB were detected with quantitative real-time PCR (qRT-PCR), Western blot and Immunofluorescence. Compared with the control group, the cell viability of HUVECs exposed to PM2.5 was significantly decreased (p < 0.05); the levels of IL-Iß and TNF-α were significantly increased, while the level of CTRP9 was significantly decreased (p < 0.05). The proportion of apoptotic cells was increased after being treated with PM2.5 (p < 0.05). Besides, the mRNA and protein levels of CD40, CD40L, IKKB, NLRP3 and caspase1 were increased comparing with the control group (p < 0.05). After CD40 silencing, the condition of inflammation and apoptosis in HUVECs exposed to PM2.5 was alleviated, and the expression levels of CD40L, IKKB, NLRP3 and caspase1 were significantly decreased (p < 0.05). Furthermore, miR-145-5p was significantly down-regulated after exposure to 200µg/mL PM2.5 (p < 0.05). After over-expression of miR-145-5p, the expression level of CD40 was decreased (p < 0.05). Taken together, PM2.5 can cause inflammation and apoptosis of HUVECs via the activation of CD40, which can be regulated by miR-145-5p. Over-expression of miR-145-5p can down-regulate CD40, further inhibiting the inflammation and apoptosis of HUVECs induced by PM2.5.
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Apoptosis/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , MicroARNs/genética , Material Particulado/toxicidad , Ligando de CD40/genética , Supervivencia Celular/efectos de los fármacos , Regulación hacia Abajo/efectos de los fármacos , Silenciador del Gen , Células Endoteliales de la Vena Umbilical Humana/patología , Humanos , Inflamación/inducido químicamente , Inflamación/patología , ARN Mensajero/metabolismo , ARN Interferente Pequeño/administración & dosificaciónRESUMEN
Cell-based therapies are recognized as the next frontier in medicine, but the translation of many promising technologies into the clinic is currently limited by a lack of remote-control inducers that are safe and can be tightly regulated. Here, we developed therapeutically active engineered cells regulated by a control system that is responsive to protocatechuic acid (PCA), a metabolite found in green tea. We constructed multiple genetic control technologies that could toggle a PCA-responsive ON/OFF switch based on a transcriptional repressor from Streptomyces coelicolor We demonstrated that PCA-controlled switches can be used for guide RNA expression-mediated control of the CRISPR-Cas9 systems for gene editing and epigenetic remodeling. We showed how these technologies could be used as implantable biocomputers in live mice to perform complex logic computations that integrated signals from multiple food metabolites. Last, we used our system to treat type 1 and type 2 diabetes in mice and cynomolgus monkeys. This biocompatible and versatile food phenolic acid-controlled transgenic device opens opportunities for dynamic interventions in gene- and cell-based precision medicine.
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Diabetes Mellitus Tipo 1/tratamiento farmacológico , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Hidroxibenzoatos/uso terapéutico , Té/química , Administración Oral , Animales , Diabetes Mellitus Tipo 1/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Femenino , Células HEK293 , Haplorrinos , Humanos , Hidroxibenzoatos/administración & dosificación , Hidroxibenzoatos/química , Masculino , Ratones , Placenta/efectos de los fármacos , Placenta/metabolismo , Embarazo , Biología SintéticaRESUMEN
Nilaparvata lugens (Stål) (Hemiptera:Delphacidae) is a serious migratory pest of rice in Asia. Pest resurgence often occurs because of insecticide overuse. Using both susceptible (TN1) and moderately resistant (Xieyou 963) cultivars, we studied the effect of foliar insecticides on the percentage of brachypterous adults, female reproductive rate, and crude fat and soluble sugar contents in third- and fifth-instar nymphs and adults. The percentage of brachypterous adults and reproductive rate of adult females developed from nymphs that fed on insecticide-treated plants varied significantly with rice cultivar, type of insecticide, and its concentration. Feeding on susceptible plants increased the percentage of brachypterous adults and reproductive rate of adult females. Also, deltamethrin increased brachypterous production relative to imidacloprid and triazophos. The highest reproductive rate was on plants treated with triazophos. All insecticide treatments in both cultivars resulted in increase of soluble sugar contents in third- and fifth-instar nymphs and adults developed from nymphs feeding on insecticide-treated rice plants. This effect was stronger on the susceptible cultivar. Changes of crude fat content after N. lugens feeding on insecticide-treated plants were related to its feeding duration. Crude fat content in adult developed from nymphs feeding on treated plants was significantly higher that on control plants. These studies showed that plant and insecticide influences on physiological ecology of this planthopper will influence its population dynamics under commercial production of rice in Asia.
Asunto(s)
Hemípteros/efectos de los fármacos , Hemípteros/crecimiento & desarrollo , Insecticidas/farmacología , Oryza/metabolismo , Análisis de Varianza , Animales , Carbohidratos/análisis , Grasas/análisis , Femenino , Hemípteros/química , Ninfa/química , Densidad de Población , Reproducción/efectos de los fármacosRESUMEN
Nilaparvata lugens (Stål) is a typical vascular feeder, primarily sucking the phloem sap of host plants. Its feeding on rice, Oryza sativa L., plants changes the pattern of allocation of assimilates between roots and shoots, and the root:shoot (R/S) ratio of assimilates is often measured as an index of physiological responses to N. lugens. The current study investigated changes in the R/S ratio of biomass, sucrose, and soluble sugar contents of rice plants in a susceptible variety (TN1) and a resistant variety (Xieyou 963). The results demonstrated that root and shoot biomasses in the two varieties linearly decreased with the increase of N. lugens infestation density. However, the relationship between changes in the R/ S ratio ofbiomass and N. lugens density differed between rice varieties, with the R/S increasing with infestation density in TN1 and decreasing in Xieyou 963. Sucrose and soluble sugar contents and their R/S values were also significantly different between the two varieties. Compared with the control that was not infested by N. lugens, the R/S values of sucrose and soluble sugar at 3 days after infestation (DAI) increased but decreased at 6 DAI in TN1. The R/S values of sucrose and soluble sugar were higher at 6 DAI than those at 3 DAI in TN1, whereas these values were lower at 6 DAI than at 3 DAI in Xieyou 963. These contrasting results suggest that physiological responses to N. lugens infestation differ between the susceptible and tolerant rice varieties.