Reactive oxygen species (ROS) modulate nitrogen signaling using temporal transcriptome analysis in foxtail millet.

Reactive oxygen species (ROS) is a chemically reactive chemical substance containing oxygen and a natural by-product of normal oxygen metabolism. Excessive ROS affect the growth process of crops, which will lead to the decrease of yield. Nitrogen, as a critical nutrient element in plants and plays a vital role in plant growth and crop production. Nitrate is the primary nitrogen source available to plants in agricultural soil and various natural environments. However, the molecular mechanism of ROS-nitrate crosstalk is still unclear. In this study, we used the foxtail millet (Setaria italica L.) as the material to figure it out. Here, we show that excessive NaCl inhibits nitrate-promoted plant growth and nitrogen use efficiency (NUE). NaCl induces ROS accumulation in roots, and ROS inhibits nitrate-induced gene expression in a short time. Surprisingly, low concentration ROS slight promotes and high concentration of ROS inhibits foxtail millet growth under long-term H2O2 treatment. These results may open a new perspective for further exploration of ROS-nitrate signaling pathway in plants.

Identification of systemic nitrogen signaling in foxtail millet (Setaria italica) roots based on split-root system and transcriptome analysis.

KEY MESSAGE: The study established split-root system (SRS) in foxtail millet, and identified the molecular regulatory mechanisms and metabolic pathways related to systemic nitrogen signaling based on this system and transcriptome analysis. The growth of crops is primarily constrained by the availability of nitrogen (N), an essential nutrient. Foxtail millet (Setaria italica L.) is a significant orphan crop known for its strong tolerance to barren conditions. Despite this, the signaling pathway of nitrogen in foxtail millet remains largely unexplored. Identifying the candidate genes responsible for nitrogen response in foxtail millet is crucial for enhancing its agricultural productivity. This study utilized the split-root system (SRS) in foxtail millet to uncover genes associated with Systemic Nitrogen Signaling (SNS). Transcriptome analysis of the SRS revealed 2158 differentially expressed genes (DEGs) implicated in SNS, including those involved in cytokinin synthesis, transcription factors, E3 ubiquitin ligase, and ROS metabolism. Silencing of SiIPT5 and SiATL31 genes through RNAi in transgenic plants resulted in reduced SNS response, indicating their role in the nitrogen signaling pathway of foxtail millet. Furthermore, the induction of ROS metabolism-related genes in response to KNO3 of the split-root System (Sp.KNO3) suggests a potential involvement of ROS signaling in the SNS of foxtail millet. Overall, this study sheds light on the molecular regulatory mechanisms and metabolic pathways of foxtail millet in relation to SNS.

High Overexpression of SiAAP9 Leads to Growth Inhibition and Protein Ectopic Localization in Transgenic Arabidopsis.

Amino acid permeases (AAPs) transporters are crucial for the long-distance transport of amino acids in plants, from source to sink. While Arabidopsis and rice have been extensively studied, research on foxtail millet is limited. This study identified two transcripts of SiAAP9, both of which were induced by NO3- and showed similar expression patterns. The overexpression of SiAAP9L and SiAAP9S in Arabidopsis inhibited plant growth and seed size, although SiAAP9 was found to transport more amino acids into seeds. Furthermore, SiAAP9-OX transgenic Arabidopsis showed increased tolerance to high concentrations of glutamate (Glu) and histidine (His). The high overexpression level of SiAAP9 suggested its protein was not only located on the plasma membrane but potentially on other organelles, as well. Interestingly, sequence deletion reduced SiAAP9's sensitivity to Brefeldin A (BFA), and SiAAP9 had ectopic localization on the endoplasmic reticulum (ER). Protoplast amino acid uptake experiments indicated that SiAAP9 enhanced Glu transport into foxtail millet cells. Overall, the two transcripts of SiAAP9 have similar functions, but SiAAP9L shows a higher colocalization with BFA compartments compared to SiAAP9S. Our research identifies a potential candidate gene for enhancing the nutritional quality of foxtail millet through breeding.

Vehicle identification using deep learning for expressway monitoring based on ultra-weak FBG arrays.

A deep learning with knowledge distillation scheme for lateral lane-level vehicle identification based on ultra-weak fiber Bragg grating (UWFBG) arrays is proposed. Firstly, the UWFBG arrays are laid underground in each expressway lane to obtain the vibration signals of vehicles. Then, three types of vehicle vibration signals (the vibration signal of a single vehicle, the accompanying vibration signal, and the vibration signal of laterally adjacent vehicles) are separately extracted by density-based spatial clustering of applications with noise (DBSCAN) to produce a sample library. Finally, a teacher model is designed with a residual neural network (ResNet) connected to a long short-term memory (LSTM), and a student model consisting of only one LSTM layer is trained by knowledge distillation (KD) to satisfy the real-time monitoring with high accuracy. Experimental demonstration verifies that the average identification rate of the student model with KD is 95% with good real-time capability. By comparison tests with other models, the proposed scheme shows a solid performance in the integrated evaluation for vehicle identification.

Crystalline Metallo-Organic Cage by Triphenylene-Cored Hexaterpyridine for Fast Iodine Capture.

In recent years, radioactive iodine capture has played an important role in nuclear waste treatment. However, most of the adsorbents possess low economic efficiency and undesirable reutilization in practical application. In this work, a terpyridine-based porous metallo-organic cage was assembled for iodine adsorption. Through synchrotron X-ray analysis, the metallo-cage was found to have a porous hierarchical packing mode with inherent cavity and packing channel. By taking advantage of polycyclic aromatic units and charged ⟨tpy-Zn2+-tpy⟩ (tpy = terpyridine) coordination sites in the structure, this nanocage exhibits an excellent ability to capture iodine in both the gas phase and aqueous medium, and the crystal state of the nanocage shows an ultrafast kinetic process of capturing I2 in aqueous solution within 5 min. The calculated maximum sorption capacities for I2 based on the Langmuir isotherm models are 1731 and 1487 mg g-1 for amorphous and crystalline nanocages, which is noticeably higher than most of the reported iodine sorbent materials in the aqueous phase. This work not only provides a rare example of iodine adsorption by a terpyridyl-based porous cage but also expands the applications of terpyridine coordination systems into iodine capture.

A semi-parametric approach for time-dependent ROC curves with nonignorable missing biomarker.

The main purpose of this paper is to survey the statistical inference for covariate-specific time-dependent receiver operating characteristic (ROC) curves with nonignorable missing continuous biomarker values. To construct time-dependent ROC curves, we consider a joint model which assumes that the failure time depends on the continuous biomarker and the covariates through a Cox proportional hazards model and that the continuous biomarker depends on the covariates through a semiparametric location model. Assuming a purely parametric model on the propensity score, we utilize instrumental variables to deal with the identifiable issue and estimate the unknown parameters of the propensity score by a simple and efficient method. In addition, when the propensity score is estimated, we develop HT and AIPW approaches to estimate our interested quantities. In the presence of nonignorable missing biomarker, our AIPW estimators of the interested quantities are still doubly robust when the true propensity score is a special parametric logistic model. At last, simulation studies are conducted to assess the performance of our proposed approaches, and a real data analysis is also carried out to illustrate its application.

Genome-Wide Identification and Expression Profiling of Cytochrome P450 Monooxygenase Superfamily in Foxtail Millet.

The cytochrome P450 monooxygenases (CYP450) are the largest enzyme family in plant metabolism and widely involved in the biosynthesis of primary and secondary metabolites. Foxtail millet (Setaria italica (L.) P. Beauv) can respond to abiotic stress through a highly complex polygene regulatory network, in which the SiCYP450 family is also involved. Although the CYP450 superfamily has been systematically studied in a few species, the research on the CYP450 superfamily in foxtail millet has not been completed. In this study, three hundred and thirty-one SiCYP450 genes were identified in the foxtail millet genome by bioinformatics methods, which were divided into four groups, including forty-six subgroups. One hundred and sixteen genes were distributed in thirty-three tandem duplicated gene clusters. Chromosome mapping showed that SiCYP450 was distributed on seven chromosomes. In the SiCYP450 family of foxtail millet, 20 conserved motifs were identified. Cis-acting elements in the promoter region of SiCYP450 genes showed that hormone response elements were found in all SiCYP450 genes. Of the three hundred and thirty-one SiCYP450 genes, nine genes were colinear with the Arabidopsis thaliana genes. Two hundred SiCYP450 genes were colinear with the Setaria viridis genes, including two hundred and forty-five gene duplication events. The expression profiles of SiCYP450 genes in different organs and developmental stages showed that SiCYP450 was preferentially expressed in specific tissues, and many tissue-specific genes were identified, such as SiCYP75B6, SiCYP96A7, SiCYP71A55, SiCYP71A61, and SiCYP71A62 in the root, SiCYP78A1 and SiCYP94D9 in leaves, and SiCYP78A6 in the ear. The RT-PCR data showed that SiCYP450 could respond to abiotic stresses, ABA, and herbicides in foxtail millet. Among them, the expression levels of SiCYP709B4, SiCYP71A11, SiCYP71A14, SiCYP78A1, SiCYP94C3, and SiCYP94C4 were significantly increased under the treatment of mesotrione, florasulam, nicosulfuron, fluroxypyr, and sethoxydim, indicating that the same gene might respond to multiple herbicides. The results of this study will help reveal the biological functions of the SiCYP450 family in development regulation and stress response and provide a basis for molecular breeding of foxtail millet.

Transcriptome Analysis Reveals Brassinolide Signaling Pathway Control of Foxtail Millet Seedling Starch and Sucrose Metabolism under Freezing Stress, with Implications for Growth and Development.

Low-temperature stress limits the growth and development of foxtail millet. Freezing stress caused by sudden temperature drops, such as late-spring coldness, often occurs in the seedling stage of foxtail millet. However, the ability and coping strategies of foxtail millet to cope with such stress are not clear. In the present study, we analyzed the self-regulatory mechanisms of freezing stress in foxtail millet. We conducted a physiological study on foxtail millet leaves at -4 °C for seven different durations (0, 2, 4, 6, 8, 10, and 12 h). Longer freezing time increased cell-membrane damage, relative conductance, and malondialdehyde content. This led to osmotic stress in the leaves, which triggered an increase in free proline, soluble sugar, and soluble protein contents. The increases in these substances helped to reduce the damage caused by stress. The activities of superoxide dismutase, peroxidase, and catalase increased reactive oxygen species (ROS) content. The optimal time point for the response to freezing stress was 8 h after exposure. The transcriptome analysis of samples held for 8 h at -4 °C revealed 6862 differentially expressed genes (DEGs), among which the majority are implicated in various pathways, including the starch and sucrose metabolic pathways, antioxidant enzyme pathways, brassinolide (BR) signaling pathway, and transcription factors, according to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment. We investigated possible crosstalk between BR signals and other pathways and found that BR signaling molecules were induced in response to freezing stress. The beta-amylase (BAM) starch hydrolase signal was enhanced by the BR signal, resulting in the accelerated degradation of starch and the formation of sugars, which served as emerging ROS scavengers and osmoregulators to resist freezing stress. In conclusion, crosstalk between BR signal transduction, and both starch and sucrose metabolism under freezing stress provides a new perspective for improving freezing resistance in foxtail millet.

Preparation of Heavily Doped P-Type PbSe with High Thermoelectric Performance by the NaCl Salt-Assisted Approach.

Thermoelectric (TE) technology, which can convert scrap heat into electricity, has attracted considerable attention. However, broader applications of TE are hindered by lacking high-performance thermoelectric materials, which can be effectively progressed by regulating the carrier concentration. In this work, a series of PbSe(NaCl)x (x = 3, 3.5, 4, 4.5) samples were synthesized through the NaCl salt-assisted approach with Na+ and Cl- doped into their lattice. Both theoretical and experimental results demonstrate that manipulating the carrier concentration by adjusting the content of NaCl is conducive to upgrading the electrical transport properties of the materials. The carrier concentration elevated from 2.71 × 1019 cm-3 to 4.16 × 1019 cm-3, and the materials demonstrated a maximum power factor of 2.9 × 10-3 W m-1 K-2. Combined with an ultralow lattice thermal conductivity of 0.7 W m-1 K-1, a high thermoelectric figure of merit (ZT) with 1.26 at 690 K was attained in PbSe(NaCl)4.5. This study provides a guideline for chemical doping to improve the thermoelectric properties of PbSe further and promote its applications.

Research on the influence of indoor thermal environment and activity levels on thermal comfort in protective clothing.

With the outbreak of infectious diseases such as Corona Virus Disease 2019, medical staff work intensively in isolated plots, medical disposable protective clothing (MDPC) has poor air condition and humidity permeability, which seriously reduces the thermal comfort of medical staff. In this paper, the effect of indoor thermal environment and activity levels on thermal comfort inside MDPC was studied by experiment. Five parts of the body were measured inside MDPC and the appropriate movements were chosen to simulate different levels of labor intensity. Meanwhile, physiological parameters and subjective thermal sensation were statistically analyzed. The results showed the influence range of different indoor temperatures on the temperature and humidity inside MDPC was about 1 °C and 10 %, respectively; it indicated that the environment inside MDPC could be improved by reducing indoor temperature, that is, a cross intelligent adjustment mode was proposed. The effect of labor intensity on the temperature inside MDPC was significantly less than that of humidity. Within 20 min, the humidity changes under moderate and heavy labor intensity were even more than 10 %, and the subjective discomfort threshold of the subjects increased by nearly 50 %. Furthermore, the maximum benefit could be obtained by concentrating cooling on back, forehead, chest and upper arm. Theoretical models of working time, labor intensity, and temperature and humidity inside MDPC under different indoor temperatures and different parts were given. In addition, acceptable regions inside MDPC which were approximately parallelogram in the enthalpy-humidity chart. These conclusions could be a reference for future thermal comfort inside MDPC research.

Pyrotinib plus capecitabine for patients with human epidermal growth factor receptor 2-positive breast cancer and brain metastases (PERMEATE): a multicentre, single-arm, two-cohort, phase 2 trial.

BACKGROUND: Patients with HER2-positive metastatic breast cancer have a high risk of developing brain metastases. Efficacious treatment options are scarce. We investigated the activity and safety of pyrotinib plus capecitabine in patients with HER2-positive metastatic breast cancer and brain metastases. METHODS: We did a multicentre, single-arm, two-cohort, phase 2 trial in eight tertiary hospitals in China. Patients aged 18 years or older who had radiotherapy-naive HER2-positive brain metastases (cohort A) or progressive disease after radiotherapy (cohort B), with an Eastern Cooperative Oncology Group performance status of 0-2, received pyrotinib 400 mg orally once daily, and capecitabine 1000 mg/m2 orally twice daily for 14 days, followed by 7 days off every 3 weeks until disease progression or unacceptable toxicity. The primary endpoint was confirmed intracranial objective response rate by investigator assessment according to the Response Evaluation Criteria In Solid Tumours (version 1.1). Activity and safety were analysed in patients with at least one dose of study drug. The study is ongoing, but recruitment is complete. The study is registered with ClinicalTrials.gov, NCT03691051. FINDINGS: Between Jan 29, 2019, and July 10, 2020, we enrolled 78 women: 51 (86%) of 59 patients in cohort A and 18 (95%) of 19 patients in cohort B had previous exposure to trastuzumab. Median follow-up duration was 15·7 months (IQR 9·7-19·0). The intracranial objective response rate was 74·6% (95% CI 61·6-85·0; 44 of 59 patients) in cohort A and 42·1% (20·3-66·5; eight of 19 patients) in cohort B. The most common grade 3 or worse treatment-emergent adverse event was diarrhoea (14 [24%] in cohort A and four [21%] in cohort B). Two (3%) patients in cohort A and three (16%) in cohort B had treatment-related serious adverse events. No treatment-related deaths occurred. INTERPRETATION: To our knowledge, this is the first prospective study showing the activity and safety of pyrotinib plus capecitabine in patients with HER2-positive breast cancer and brain metastases, especially in radiotherapy-naive population. This combination deserves further validation in a randomised, controlled trial. FUNDING: National Cancer Centre Climbing Foundation Key Project of China, Jiangsu Hengrui Pharmaceuticals. TRANSLATION: For the Chinese translation of the abstract see Supplementary Materials section.

Key residues for maintaining architecture, assembly of plant hormone SA receptor NPR1.

Salicylic acid (SA) is a pivotal hormone required for the development of resistance to many pathogens in plants. As an SA receptor, NPR1(Nonexpressor of Pathogenesis-Related Genes 1) plays a key regulatory role in the plant immune response. The function of NPR1 is dependent on the alteration of its oligomer-to-monomer. Research in recent years has proven that NPRs perceive SA and regulate the expression of downstream defense genes, but the mechanism of NPR1 oligomer-to-monomer conversion remains unclear. In this paper, we mainly studied the oligomerization of NPR1. By mutation experiments on some residues in the BTB domain involved in protein interactions, we found that the residue His80 plays a key role in the oligomerization of NPR1. We also found that NPR1, interacting with zinc ions at a ratio close to 1:1, was independent of the residue His80. These findings may help us to understand the conformational conversion of NPR1.

Intrusion identification using GMM-HMM for perimeter monitoring based on ultra-weak FBG arrays.

Intrusion identification has been an intractable task for perimeter security. One of the primary challenges is to possess high identification rate over a long-distance range monitoring. This paper proposes an intrusion identification scheme based on ultra-weak fiber Bragg grating (UWFBG) arrays. The scheme is acquired by the combination of a Gaussian mixture model (GMM) and a hidden Markov model (HMM). The time dependencies are obtained by the analysis of relevant sensors in UWFBG arrays from the procedure of intrusions. The features extracted from vibration signals with time dependencies are used as the input of GMM-HMM. The GMM-HMM simultaneously analyzes features and time dependencies to identify intrusion. Experimental demonstration verifies that the proposed scheme can identify three intrusions (walking, knocking and climbing) and two non-intrusions (heavy truck passing and wind blowing) with the average identification rate of 98.2%. By the comparison tests with other six classifiers, the proposed GMM-HMM scheme shows a solid performance in the integrated evaluation for intrusion identification.

3D Cationic Polymeric Network Nanotrap for Efficient Collection of Perrhenate Anion from Wastewater.

Rhenium is one of the most valuable elements found in nature, and its capture and recycle are highly desirable for resource recovery. However, the effective and efficient collection of this material from industrial waste remains quite challenging. Herein, a tetraphenylmethane-based cationic polymeric network (CPN-tpm) nanotrap is designed, synthesized, and evaluated for ReO4- recovery. 3D building units are used to construct imidazolium salt-based polymers with positive charges, which yields a record maximum uptake capacity of 1133 mg g-1 for ReO4- collection as well as fast kinetics ReO4- uptake. The sorption equilibrium is reached within 20 min and a kd value of 8.5 × 105 mL g-1 is obtained. The sorption capacity of CPN-tpm remains stable over a wide range of pH values and the removal efficiency exceeds 60% for pH levels below 2. Moreover, CPN-tpm exhibits good recyclability for at least five cycles of the sorption-desorption process. This work provides a new route for constructing a kind of new high-performance polymeric material for rhenium recovery and rhenium-contained industrial wastewater treatment.

Jaridon 6, a new diterpene from Rabdosia rubescens (Hemsl.) Hara, can display anti-gastric cancer resistance by inhibiting SIRT1 and inducing autophagy.

Tumor resistance is the main cause of treatment failure and is associated with many tumor factors. Jaridon 6, a new diterpene extracted from Rabdosia rubescens (Hemsl.) Hara, which has been previously extracted by our research team, has been tested having more obvious advantages in resistant tumor cells. However, its mechanism is unclear. In this study, we studied the effect and the specific mechanism of Jaridon 6 in resistant gastric cancer cells. Cytotoxicity test, colony test, western blotting, and nude test verified the anti-drug resistance ability of Jaridon 6 in the MGC803/PTX and MGC803/5-Fu cells. Jaridon 6 has shown obvious inhibitory effects in the sirtuin 1 (SIRT1) enzyme test. Transmission electron microscopy and immunofluorescence tests further proved the autophagic action of Jaridon 6. Jaridon 6 could inhibit the proliferation of the resistant gastric cancer cell in vivo and in vitro. Jaridon 6 inhibited SIRT1 enzyme and induced autophagy by inhibiting the phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) pathway. Thus, it may be considered for treating gastric cancer resistance by individual or combined administration, as an SIRT1 inhibitor and autophagy inducer.

Sulfur application reduces cadmium uptake in edible parts of pakchoi (Brassica chinensis L.) by cadmium chelation and vacuolar sequestration.

Sulfur (S) application in pakchoi (Brassica chinensis L.) cultivation is vital for reducing cadmium (Cd) accumulation in the plants. However, the mechanism of S application on Cd uptake and translocation in pakchoi is unclear. In this study, a hydroponic experiment was performed to investigate the effects of S application on Cd accumulation in pakchoi at one Cd concentration (50 µM, in comparison to the control condition, 0 µM) and three S levels (0, 2, 4 mM). The results showed that excessive S application (4 mM) reduced Cd accumulation and alleviated pakchoi growth inhibition caused by Cd stress in shoots and roots. With increased S application, the proportion of Cd in the vacuolar fraction and the proportion of NaCl-extractable Cd increased in roots. Additionally, S application increased the content of glutathione (GSH) and phytochelatins (PCs). The reduced Cd uptake and accumulation in pakchoi shoots could have been due to increased Cd chelation and vacuolar sequestration in roots. In addition, sufficient S application (2 mM) increased the expression of γ-glutamylcysteine synthetase (GSH1) and nicotinamide synthase (NAS) in roots, and excessive S application upregulated the expression of ATP sulfurylase (ATPS) and phytochelatin synthase (PCs). This study provides evidence for the mechanism of mitigating Cd toxicity in pakchoi and will be helpful for developing strategies to reduce Cd accumulation in the edible parts of pakchoi through S fertilizer application.

A Wheat WRKY Transcription Factor TaWRKY46 Enhances Tolerance to Osmotic Stress in transgenic Arabidopsis Plants.

WRKY transcription factors play central roles in developmental processes and stress responses of wheat. Most WRKY proteins of the same group (Group III) have a similar function in abiotic stress responses in plants. TaWRKY46, a member of Group III, was up-regulated by PEG treatment. TaWRKY46-GFP fusion proteins localize to the nucleus in wheat mesophyll protoplasts. Overexpression of TaWRKY46 enhanced osmotic stress tolerance in transgenic Arabidopsis thaliana plants, which was mainly demonstrated by transgenic Arabidopsis plants forming higher germination rate and longer root length on 1/2 Murashige and Skoog (MS) medium containing mannitol. Furthermore, the expression of several stress-related genes (P5CS1, RD29B, DREB2A, ABF3, CBF2, and CBF3) was significantly increased in TaWRKY46-overexpressing transgenic Arabidopsis plants after mannitol treatment. Taken together, these findings proposed that TaWRKY46 possesses vital functions in improving drought tolerance through ABA-dependent and ABA-independent pathways when plants are exposed to adverse osmotic conditions. TaWRKY46 can be taken as a candidate gene for transgenic breeding against osmotic stress in wheat. It can further complement and improve the information of the WRKY family members of Group III.

Oroxyloside inhibits angiogenesis through suppressing internalization of VEGFR2/Flk-1 in endothelial cells.

Increasing flavonoids have been reported to possess anti-angiogenic effects. Inhibition of angiogenesis plays a critical role in the treatment of cancer, especially in advanced metastatic cancer. In this study, we assessed the effect of Oroxylin A-7-glucuronide (Oroxyloside), a main metabolite of Oroxylin A, on angiogenesis in human endothelial cell-like EA.hy926 cells. Oroxyloside suppressed the migration and tube formation of EA.hy926 cells. Meanwhile, microvessels sprouting from aortic rings and new blood vessels on the chicken chorioallantoic membrane (CAM) were also inhibited. Mechanism studies showed that Oroxyloside reduced the autophosphorylation of vascular endothelial growth factor receptor 2 (VEGFR2/Flk-1) while it up-regulated the expression of R-Ras and VE-cadherin. In consequence, Oroxyloside inhibited the downstream Akt/MAPK/NF-κB pathways and then decreased the nuclear translocation and DNA binding ability of NF-κB. Furthermore, in vivo study showed that Oroxyloside exhibited a potential anti-angiogenic effect in Matrigel plug assay and inhibited growth of xenografted tumors with low systemic toxicity, which could be ascribed to the inhibition of VEGFR2 internalization. Taken together, these results suggested that Oroxyloside could inhibit angiogenesis in vitro and in vivo via suppressing the internalization of VEGFR2 and might serve as a potential antitumor agent.

Generation and characterization of a paclitaxel-resistant human gastric carcinoma cell line.

The main aim of this study was to establish a novel paclitaxel (PTX)-resistant human gastric carcinoma cell line and to investigate its biological significance. A cell line, MGC803/PTX, was established by gradually increasing PTX density on the basis of MGC803 over a period of 10 months. In addition, a pair of resistant cell lines (SW620 and SW620/PTX) were added to further explain the resistant mechanism of PTX. The drug resistance index and stability of MGC803/PTX cells were detected using the Cell Counting Kit-8 method. The morphological features were observed using inverted microscopy. Apoptosis was measured by flow cytometry (FCM) and Hoechst 33258 fluorescence staining. The distribution of the cell cycle was determined by FCM, and protein expressions of P-gp, Bcl-2, Bax, and PARP were detected by western blot analysis. When characterizing the resistance in vitro, we found that MGC803/PTX cells were 10.3-fold more resistant to PTX compared with MGC803 cells. In addition, MGC803/PTX cells showed cross-resistance to 5-fluorouracil and adriamycin. FCM and Hoechst 33258 fluorescence staining indicated that MGC803/PTX cells had a significantly lower percentage of apoptotic cells after treatment with PTX compared with MGC803 cells. Other differences between parental cells and resistant cells included morphology, proliferation rate, doubling time, cell cycle distribution, and colony-formation rate. Western blot analysis indicated that P-gp, Bcl-2, and PARP protein were more abundant in MGC803/PTX and SW620/PTX cells compared with MGC803 and SW620 cells, whereas Bax protein levels were lower in resistant cells. Furthermore, MGC803/PTX cells showed obvious resistance to PTX in vivo. To our knowledge, this is the first report on the establishment of a PTX-resistant MGC803 cell line, which is an important tool to explore the resistance of anticancer drugs and to overcome tumor drug resistance.

Effects of Continuous Phase and Crosslinking Agent on the Rheological Behaviors and Properties of Cationic Poly(urethane-acrylate) Emulsifier-Free Microemulsions.

Cationic poly(urethane-acrylate) emulsifier-free microemulsions are prepared by seeded emulsion polymerization (SPUA) or in-situ emulsion polymerization (IPUA) with (a) or without (b) crosslinking agent hydroxyethyl acrylate (HEA). Furthermore, no surfactant is introduced into the reaction system. The effect of continuous phase and HEA on the rheological behavior and properties of microemulsions are studied. The results show that IPUA using vinylmonomers as the continuous phase and with HEA (IPUAMB-b) has the highest ζ value, lowest coagulation ratio, smallest particle size of 48 nm and apparent core-shell structures. Rheological research has concluded that the zero-shear viscosity of IPUAB-b and IPUAM-b emulsion is 6.03×102 and 7.89 Pa·s, respectively, and presents pseudoplastic behavior. However, IPUAMB-b and SPUA-b display Newtonian behavior with η0 of 0.136 Pa·s and 0.0823 Pa·s, respectively. PUA emulsions display similar rheological behavior when NMP is replaced by the mixture of MMA and BA.