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1.
Med Sci Monit ; 26: e921233, 2020 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-32032347

RESUMO

BACKGROUND Osteosarcoma is a common malignant tumor of musculoskeletal stromal cells. Osteosarcoma clinical behavior depends mostly on the histologic grade, the site of primary tumor, the response to chemotherapy, and the presence of pulmonary metastases. The aim of this study was to knockout SHOX CNE9/10 in U2OS osteosarcoma cells and to analyze the effects on cell growth and apoptosis. MATERIAL AND METHODS U2OS cells with CNE9 knockout and U2OS cells with CNE10 knockout were established via the CRISPR/Cas9 system. Sanger sequencing was used to detect the success of the knockdown experiment. Western blotting and quantitative polymerase chain reaction were used to detect the expression levels of short stature homeobox-containing gene (SHOX) protein and messenger RNA (mRNA) after knockdown of CNE9 and CNE10. The cell viability and apoptotic rate were detected by the Cell Counting Kit-8 method and by flow cytometry. RESULTS The Sanger sequencing results showed that the knockdown experiment was successful. The levels of SHOX mRNA and protein were significantly reduced after knocking down CNE9 and CNE10. Knockdown of CNE9 and CNE10 significantly increased the growth and inhibited the apoptosis of U2OS osteosarcoma cells. CNE9/CNE10 knockdown U2OS cells were successfully constructed. CONCLUSIONS Knockdown of CNE9 and CNE10 promoted U2OS cell growth and inhibited apoptosis by decreasing SHOX expression. This CNE9/CNE10 knockout U2OS cell model could provide a bridge for the research on SHOX and CNEs in osteosarcoma.

2.
Int J Biol Macromol ; 2020 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-32035959

RESUMO

ß-Amyloid (Aß) plays an important role in the pathogenesis of Alzheimer's disease (AD). However, there is still no effective Aß-targeting drugs for AD treatment. In this study, we explored the effect and mechanism of Sodium Tanshinone IIA Sulfonate (STS) on AD. Aß-treated HT22 cells, an immortalized mouse hippocampal neuronal cell line, were employed. Different dosages of STS (0.1, 1 and 10 µM) were selected. STS improved cell viability and protected against Aß-induced apoptosis in a dose-dependent manner. Furthermore, the levels of reactive oxygen species (ROS) and malondialdehyde (MDA) were decreased, while the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were significantly increased after STS treatment. STS decreased the levels of phosphorylate PKR-like (p-PERK), phosphorylate eukaryotic initiation factor 2 (p-eIF2α), phosphorylate inositol-requiring enzyme (p-IRE1α), X-box binding protein 1 (XBP1) and binding immunoglobulin heavy chain protein (Bip), while increased protein disulfide isomerase (PDI) levels in Aß-treated HT22 cells. In addition, the levels of insulin degrading enzymes (IDE) and Nepterrilysin (NEP) (or call it CD10) were significantly increased after STS treatment. Taken together, these results indicated that STS might be effective in treating AD via increasing the levels of Aß-degrading enzymes.

3.
Expert Opin Drug Discov ; : 1-14, 2020 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-32000537

RESUMO

Introduction: Duchenne muscular dystrophy (DMD) is an X-linked handicapping disease due to the loss of an essential muscle protein dystrophin. Dystrophin-null animals have been extensively used to study disease mechanisms and to develop experimental therapeutics. Despite decades of research, however, treatment options for DMD remain very limited.Areas covered: High-throughput high-content screening and precision medicine offer exciting new opportunities. Here, the authors review animal models that are suitable for these studies.Expert opinion: Nonmammalian models (worm, fruit fly, and zebrafish) are particularly attractive for cost-effective large-scale drug screening. Several promising lead compounds have been discovered using these models. Precision medicine for DMD aims at developing mutation-specific therapies such as exon-skipping and genome editing. To meet these needs, models with patient-like mutations have been established in different species. Models that harbor hotspot mutations are very attractive because the drugs developed in these models can bring mutation-specific therapies to a large population of patients. Humanized hDMD mice carry the entire human dystrophin gene in the mouse genome. Reagents developed in the hDMD mouse-based models are directly translatable to human patients.

4.
EBioMedicine ; 52: 102638, 2020 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-32014820

RESUMO

BACKGROUND: To improve the early diagnosis of hepatocellular carcinoma (HCC), more effective diagnostic biomarkers are needed. A combination of biomarkers is reported to distinguish individuals with early-stage HCC from at-risk individuals. METHODS: Participants in this study were recruited from six hospitals in China. Literature review was used to choose 19 candidate proteins, a case-control study in the discovery stage was used to identify five proteins (P5) that constituted a diagnostic model. In the training and validation stages, the effectiveness of P5 for detecting early-stage HCC was tested (cross-sectional study). Finally, a nested case-control study independent of the other stages was set up to evaluate the P5 in the preclinical diagnosis of HCC. FINDINGS: Between February 2013 and June 2017, a total of 1396 participants were recruited. A panel of 5 proteins (P5: OPN, GDF15, NSE, TRAP5 and OPG) showed high diagnostic accuracy when differentiating the early-stage HCC from the at-risk group, with AUCs of 0·892, 0·907 and 0·852 for the training stage, validation cohort 1 and cohort 2 data sets, respectively. In the prediction set, the sensitivity of P5 for diagnosing preclinical HCC increased with time, starting from 12 months before to the time of definitive clinical diagnosis (range, 46·15% to 86·67%). INTERPRETATION: The P5 panel has the potential to screen populations at high risk of developing HCC and can enable the early diagnosis of HCC. FUNDING: Research supported by grants from eight funds. All sources of funding were declared at the end of the text.

5.
Colloids Surf B Biointerfaces ; 189: 110831, 2020 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-32058252

RESUMO

Stent intervention as available method in clinic has been widely applied for cardiovascular disease treatment for decades. However, the restenosis caused by late thrombosis and hyperplasia still limits the stents long-term application, and the essential cause is usually recognized as endothelial functionalization insufficiency of the stent material surface. Here, we address this limitation by developing a pro-endothelial-functionalization surface that immobilized a natural factors-loaded nanoparticle, exosome, onto the poly-dopamine (PDA) coated materials via electrostatic binding. This PDA/Exosome surface not only increased the endothelial cells number on the materials, but also improved their endothelial function, including platelet endothelial cell adhesion molecule-1 (CD31) expression, cell migration and nitric oxide release. The pro-inflammation macrophage (M1 phenotype) attachment and synthetic smooth muscle cell proliferation as the interference factors for the endothelialization were not only inhibited by the PDA/Exosome coating, while the cells were also regulated to anti-inflammation macrophage (M2 phenotype) and contractile smooth muscle cell, which may contribute to endothelialization. Thus, it can be summarized this method has potential application on surface modification of cardiovascular biomaterials.

6.
Artigo em Inglês | MEDLINE | ID: mdl-32053348

RESUMO

The Janus amphiphilic particles have gained much attention for their important application value in areas as diverse asinterfacial modification, sensors, drug delivery, optics and actuators. In this work, we prepared Janus amphiphilic nanosheets composed of nitrogen doped stratiform meso-macroporous carbons (NMC) and molybdenum sulfide (MoS2) for hydrophilic and hydrophobic side, respectively. The dicyandiamide and glucose were used as precursors for synthesizing two-dimensional nitrogen doped meso-macroporous carbons and the molybdate could be anchored by the functional groups, on the surface of carbon layers, then transform into uniformly MoS2 to form the Janus amphiphilic layer by layer NMC/MoS2 support. Transmission electron microscope (TEM),Scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared (FTIR) spectroscopy, are used to demonstrate the successful preparation of Janus materials. As typical interfacial enzyme, Candida Rugosa lipase (CRL) immobilized on the Janus amphiphilic NMC/MoS2 support brought forth to improvement of its performance due to the Janus nanosheets can be easily attached on the oil-aqueous interface for better catalytic activity (interfacial activation of lipases). The obtained immobilized lipase (NMC/MoS2@CRL) exhibited satisfactory lipase-loading (193.1 mg protein per g), specific hydrolytic activity (95.76 U g-1), thermostability (at 55 °C, 84 % of the initial activity remained after 210 min), pH-flexibility and recyclability (60 % of the initial activity remained after 9 runs). In terms of its application, the esterification rate of using NMC/MoS2@CRL (75 %) is higher than NMC@CRL (20 %), MoS2@CRL (11.8 %) in "oil-water" biphase and CRL as well as NMC/MoS2@CRL in one-phase. Comparing with the free CRL, NMC@CRL and MoS2@CRL, the Janus amphiphilic NMC/MoS2 served as carrier exhibited more optimal performance and practicability.

8.
Epileptic Disord ; 2020 Feb 11.
Artigo em Inglês | MEDLINE | ID: mdl-32051108

RESUMO

Early infantile epileptic encephalopathy type 28 is a refractory epilepsy with early onset, poor prognosis, and hereditary causes. WW domain-containing oxidoreductase (WWOX) gene mutation can result in epileptic encephalopathy, but the mechanism remains unclear. We present the case of a patient with epilepsy and WWOX compound heterozygous mutations. The seizures manifested as tonic-clonic, convulsive and were refractory to drugs. Magnetic resonance imaging showed a widened subarachnoid space and thin corpus callosum. The patient died from asphyxia at the age of one year and 23 days. Peripheral blood was taken from the patient and his parents, and whole-exome sequencing was investigated to determine possible gene mutation. Two compound heterozygous mutations were identified: c.172+1G>C (with no amino acid change) and c.984C>G (amino acid change: p.Tyr328Ter). The pathophysiology of epileptic encephalopathy related to the WWOX gene remains to be determined, and further studies are required to elucidate possible mechanisms.

9.
ACS Appl Mater Interfaces ; 12(2): 2671-2678, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31899615

RESUMO

Transition-metal sulfides have been considered as promising anode materials for lithium-ion batteries (LIBs) due to their high theoretical specific capacity and superior electrochemical performance. However, the large volume change during the discharge/charge process causes structural pulverization, resulting in rapid capacity decline and the loss of active materials. Herein, we report Co1-xS hollow spheres formed by in situ growth on reduced graphene oxide layers. When evaluated as an anode material for LIBs, it delivers a specific capacity of 969.8 mAh·g-1 with a high Coulombic efficiency of 96.49% after 90 cycles. Furthermore, a high reversible capacity of 527.2 mAh·g-1 after the 107th cycle at a current density of 2.5 A g-1 is still achieved. The results illustrate that in situ growth on the graphene layers can enhance conductivity and restrain volume expansion of cobalt sulfide compared with ex situ growth.

10.
Nucleic Acids Res ; 2020 Jan 16.
Artigo em Inglês | MEDLINE | ID: mdl-31943114

RESUMO

RNA aptamers that bind HIV-1 reverse transcriptase (RT) inhibit RT in enzymatic and viral replication assays. Some aptamers inhibit RT from only a few viral clades, while others show broad-spectrum inhibition. Biophysical determinants of recognition specificity are poorly understood. We investigated the interface between HIV-1 RT and a broad-spectrum UCAA-family aptamer. SAR and hydroxyl radical probing identified aptamer structural elements critical for inhibition and established the role of signature UCAA bulge motif in RT-aptamer interaction. HDX footprinting on RT ± aptamer shows strong contacts with both subunits, especially near the C-terminus of p51. Alanine scanning revealed decreased inhibition by the aptamer for mutants P420A, L422A and K424A. 2D proton nuclear magnetic resonance and SAXS data provided constraints on the solution structure of the aptamer and enable computational modeling of the docked complex with RT. Surprisingly, the aptamer enhanced proteolytic cleavage of precursor p66/p66 by HIV-1 protease, suggesting that it stabilizes the productive conformation to allow maturation. These results illuminate features at the RT-aptamer interface that govern recognition specificity by a broad-spectrum antiviral aptamer, and they open new possibilities for accelerating RT maturation and interfering with viral replication.

11.
Sci Rep ; 10(1): 744, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31937843

RESUMO

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

12.
J Exp Clin Cancer Res ; 39(1): 15, 2020 Jan 17.
Artigo em Inglês | MEDLINE | ID: mdl-31948467

RESUMO

In the original publication of this manuscript [1], there are three errors in Fig. 1. The identified errors do not affect the conclusions of the work.

13.
J Chem Phys ; 152(3): 034502, 2020 Jan 21.
Artigo em Inglês | MEDLINE | ID: mdl-31968977

RESUMO

We propose a microscopic theory for the decoupling of self-diffusion and structural relaxation in glass-forming liquids within the Elastically Collective Nonlinear Langevin Equation (ECNLE) activated dynamics framework. Our central hypothesis is that the heterogeneity relevant to this problem is static fluctuations of local density on the scale of 3-4 particle diameters and how this changes local packing correlations. These fluctuations modify the degree of dynamical cage expansion that mechanistically couples intracage large amplitude hopping and longer range collective elasticity in ECNLE theory. Decoupling only emerges in the deeply supercooled regime where the strongly temperature dependent elastic barrier becomes non-negligible relative to its noncooperative local analog. The theory makes predictions for various aspects of the decoupling phenomenon, including apparent fractional power law Stokes-Einstein behavior, that appear to be consistent with experiments and simulations on hard sphere fluids and molecular liquids. Of central importance is a microscopic connection between the barrier fluctuation variance and most probable barrier height. Sensible results are also obtained for the nonexponential stretching of a generic relaxation time correlation function and its temperature evolution. Nonuniversality can arise from the relative importance of the local and collective barriers (related to fragility) and the precise magnitude of the length scale that defines the transition from local cage to elastic physics. Comparison is made with a traplike model based on a Gaussian distribution of barriers.

14.
J Cell Mol Med ; 2020 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-32000299

RESUMO

Epidemiological studies have found that diabetes and cognitive dysfunction are closely related. Quercetin has been certified with the effect on improving diabetes mellitus (DM) and cognitive impairment. However, the effect and related mechanism of quercetin on diabetic encephalopathy (DE) are still ambiguous. In this study, we used the db/db mice (diabetic model) to discover whether quercetin could improve DE through the Sirtuin1/NLRP3 (NOD-, LRR- and pyrin domain-containing 3) pathway. Behavioural results (Morris water maze and new object recognition tests) showed that quercetin (70 mg/kg) improved the learning and memory. Furthermore, quercetin alleviated insulin resistance and the level of fasting blood glucose. Besides, Western blot analysis also showed that quercetin increased the protein expressions of nerve- and synapse-related protein, including postsynapticdensity 93 (PSD93), postsynapticdensity 95 (PSD95), brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in the brain of db/db mice. Quercetin also increased the protein expression of SIRT1 and decreased the expression of NLRP3 inflammation-related proteins, including NLRP3, the adaptor protein ASC and cleaved Caspase-1, the pro-inflammatory cytokines IL-1ß and IL-18. In conclusion, the present results indicate that the SIRT1/NLRP3 pathway may be a crucial mechanism for the neuroprotective effect of quercetin against DE.

15.
J Cell Mol Med ; 2020 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-31989795

RESUMO

Sodium tanshinone IIA sulfonate (STS) has been reported to prevent Alzheimer's disease (AD). However, the mechanism is still unknown. In this study, two in vitro models, Aß-treated SH-SY5Y cells and SH-SY5Y human neuroblastoma cells transfected with APPsw (SH-SY5Y-APPsw cells), were employed to investigate the neuroprotective of STS. The results revealed that pretreatment with STS (1, 10 and 100 µmol/L) for 24 hours could protect against Aß (10 µmol/L)-induced cell toxicity in a dose-dependent manner in the SH-SY5Y cells. Sodium tanshinone IIA sulfonate decreased the concentrations of reactive oxygen species, malondialdehyde, NO and iNOS, while increased the activities of superoxide dismutase and glutathione peroxidase in the SH-SY5Y cells. Sodium tanshinone IIA sulfonate decreased the levels of inflammatory factors (IL-1ß, IL-6 and TNF-α) in the SH-SY5Y cells. In addition, Western blot results revealed that the expressions of neprilysin and insulin-degrading enzyme were up-regulated in the SH-SY5Y cells after STS treatment. Furthermore, ELISA and Western blot results showed that STS could decrease the levels of Aß. ELISA and qPCR results indicated that STS could increase α-secretase (ADAM10) activity and decrease ß-secretase (BACE1) activity. In conclusion, STS could protect against Aß-induced cell damage by modulating Aß degration and generation. Sodium tanshinone IIA sulfonate could be a promising candidate for AD treatment.

16.
Anal Chem ; 92(2): 2043-2051, 2020 Jan 21.
Artigo em Inglês | MEDLINE | ID: mdl-31894964

RESUMO

Identification of metabolites at the trace level in complex samples is still one of the major challenges in untargeted metabolomics. One formula in the metabolomic database is always corresponding to more than one candidate, which increases the difficulty and cost in the subsequent process of standard compound matching. In this study, we developed an effective method for amine metabolite identification by hydrogen-deuterium scrambling (HDS) based on chemical isotope labeling coupled with liquid chromatography-mass spectrometry (HDS-CIL-LC-MS). After d4-4-(N,N-dimethylamino)phenyl isothiocyanate (d4-DMAP) labeling, the labeled amine metabolites can produce HDS under collision-induced dissociation (CID). The HDS can effectively reflect the number of labile hydrogen atoms in amine metabolites and thus distinguish amine isomers with different functional groups. The developed HDS-CIL-LC-MS method was applied to the determination of amine metabolites in mice feces, in which the amine candidates obtained by the database based on chemical formula searching were reduced by 64% on average, which greatly reduces the cost of standard compound matching. Taken together, the developed HDS-CIL-LC-MS analysis was demonstrated to be a promising method for untargeted metabolomics and a novel strategy for deciphering tandem mass spectrometry (MS2) spectra.

17.
Ying Yong Sheng Tai Xue Bao ; 31(1): 139-147, 2020 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-31957390

RESUMO

To reveal the coupling effect of water and fertilizer on the yield of spring maize under shallow-buried drip irrigation in semi-arid area of western Liaoning, a field experiment was conducted with the quadratic regression orthogonal design of three factors (water, nitrogen and potassium) crossed with five levels in 2017-2018. A quadratic regression model was established with yield (Y) as the dependent variable and irrigation amount (W), nitrogen (N) and potassium (K) application amounts as independent variables to analyze the coupling relationships between Y and W, N and K, respectively. The results showed that the shallow-drip irrigation water-fertilizer coupling had significant impact on yield. The single factor of W, N and K promoted the yield, with their effects ranking as W>N>K. The effect of two-factor interaction on yield increased first and then decreased which ranked as WN>WK>NK. Considering the three-factor coupling effect on yield, the combination of abundant water, nitrogen and potassium was the highest, followed by high water, nitrogen and potassium, and low water, nitrogen and potassium the lowest. As the optimal treatments found by the model, we obtained the suitable water-fertilizer application range of shallow-buried drip irrigation with higher target yield of 8000-8810 kg·hm-2, that was, the irrigation amount was 43-61 mm, nitrogen 138-343 kg·hm-2 and potassium 79-163 kg·hm-2 under the ambient natural rainfall. The results provided a referable basis for application of water-fertilizer integrated cropping pattern under shallow-buried drip irrigation in semi-arid area of northern China.


Assuntos
Fertilizantes , Zea mays , Irrigação Agrícola , Agricultura , Biomassa , China , Nitrogênio , Água
18.
Calcif Tissue Int ; 2020 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-31897529

RESUMO

Ossification of the ligamentum flavum (OLF) is characterized by a process of ectopic bone formation in the ligamentum flavum. The definitive pathophysiology of OLF still remains unclear, but the epigenetic m6A modification plays an important role in OLF. In addition, no studies have reported the function of ALKBH5 in OLF development. In this study, we investigated the function of the m6A demethylation enzyme ALKBH5 in OLF. To evaluate the function of ALKBH5, OLF tissues and normal ligamentum flavum tissues were collected. In vitro methods, including HE, IHC and western blotting assays, were used to evaluate the association of ALKBH5 with OLF. In addition, we verified the effects of ALKBH5 on osteogenesis using alizarin red and ALP staining. MeRIP q-PCR was performed to investigate the methylation level of BMP2. Moreover, the mechanism of ALKBH5-mediated regulation of the ossification of the ligamentum flavum cells through the AKT signaling pathway was also verified. The present study showed that the expression of ALKBH5 increased in OLF tissues. The overexpression of ALKBH5 increased the expression of osteogenic genes and promoted the ossification of ligamentum flavum cells. Furthermore, BMP2 was significantly enriched in the ligamentum flavum cells of the anti-m6A group compared with those of the IgG group. The overexpression of ALKBH5 led to the activation of p-AKT, and BMP2 was regulated by ALKBH5 through the AKT signaling pathway. ALKBH5 promoted the osteogenesis of the ligamentum flavum cells through BMP2 demethylation and AKT activation. ALKBH5 was shown to be an important demethylation enzyme in OLF development.

19.
J Craniofac Surg ; 31(1): 214-218, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31652219

RESUMO

PURPOSE: The aim of this study was to evaluate the efficacy, feasibility, and safety of the endoscopic optic canal and orbital apex decompression for patients with traumatic orbital apex syndrome. DESIGN: Retrospective, noncomparative case series. METHOD: Thirty-one patients (31 eyes) with traumatic orbital apex syndrome underwent endoscopic transethmosphenoid optic canal and orbital apex decompression at the Eye Hospital of Wenzhou Medical University from May 1st, 2012 to May 1st, 2018. In each case, the indication of surgery was that patient with traumatic orbital apex syndrome failed to respond to corticosteroids. Patients were followed up to 6 months after surgery. Best corrected visual acuity, visual field, ptosis, ophthalmoplegia, hypoesthesia, and pupil before and after surgery were compared. RESULT: All patients presented visual decline (including 5 patients with no light perception), ptosis, ophthalmoplegia, diplopia, pupil dysfunction, and visual field defect, and 20 of them also presented hypoesthesia. Nineteen of 31 (61.3%) patients gained improvement of best-corrected visual acuity after surgery, 7 of them gained 20/20 BCVA, and visual field showed improvement in 20 patients. Ptosis and ophthalmoplegia of all patients recovered in various degree; diplopia also relieved relatively. The function of the pupil was also improved in most patients (27/31, 87.1%). The improvement of hypoesthesia was also observed in most patients. No serious complications occurred. CONCLUSION: Endoscopic transethmosphenoid optic canal and orbital apex decompression seems to be a feasible, efficient, and safe approach for traumatic orbital apex syndrome patients.

20.
Spectrochim Acta A Mol Biomol Spectrosc ; 227: 117540, 2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-31680040

RESUMO

A novel and highly selective fluorescent 1,8-naphthalimide-based probe, 3, was designed and synthesized for rapid Cu2+ detection in a CH3CN-H2O (3:1, v/v, pH = 7.4) solution by means of a distinct hydrolysis mechanism via its Cu2+-promoting feature. Upon treatment with Cu2+, the fluorescence response of probe 3 at 550 nm abruptly decreased, which was visible to the naked eye, and this response was accompanied by a clear change of the color of the solution; the color changed from the original yellow color to colorless. This color change occurred due to the Cu2+-promoted hydrolysis of 3, which yielded a fluorescence-quenched product. It is inspiring that probe 3 exhibited excellent sensitivity, a short response time and strong anti-interference recognition. Compared with the allowable amount of Cu2+ (∼20 µM) in drinking water, the detection limit of 3 for Cu2+ is calculated to be 9.15 nM, which is much lower than the amount defined by standards. The probe can be successfully applied for the determination of Cu2+ in real aqueous samples. Furthermore, probe 3 can be used as a fluorescent sensor to detect Cu2+ in biological environments, demonstrating its low toxicity to organisms and good cell permeability in live cell imaging.

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