Biophysical phenotyping of single-cell based on impedance and application for individualized precision medicine.

Single-cell biophysical characterization based on impedance measurement is an advantageous approach due to its label-free, high-efficiency, cost-effective and real-time capability. Biophysical phenotyping can yield timely and rich information on physiological and pathological state of cells for disease diagnosis, drug screening, precision medicine, etc. However, precise measurement on single-cell impedance is challenging, particularly hard to figure out the detailed biophysical parameters of single cell due to coupling and complexity of impedance model. Here, we propose an analytic determination method to decode single-cell electrophysiological parameters (including cell-substrate interface capacitance, cell membrane capacitance, cell membrane conductivity, and cytoplasm conductivity) from the impedances measured at optimized frequencies by using analytic solution rather than spectrum fitting. With this simple and fast analytic solution method, the physiological parameters of single cell in natural adhesion state can be accurately determined in real time. We validate this cell parameter determination method in monitoring the change of cell adhesion under hydraulic effects and exploring electrophysiological differences among MCF-7, HeLa, Huh7, and MDA-MB-231 cell lines. Particularly, we apply the approach to optimize tumor treating fields (TTFields) therapy, realizing individualized precision medicine. Our work provides an accurate and efficient approach for characterizing single-cell biophysical properties with real-time, in-situ, label-free, and less invasive advantages.

MELK aggravates lung adenocarcinoma by regulating EZH2 ubiquitination and H3K27me3 histone methylation of LATS2.

We tried to elucidate the possible roles of maternal embryonic leucine pull chain kinase (MELK) in lung adenocarcinoma (LUAD) growth and metastasis. Differentially expressed genes in LUAD samples were analysed by the GEPIA database. Clinical tissue samples and cells were collected for MELK, EZH2 and LATS2 expression determination. Co-IP assay was used to verify the interaction between EZH2 and MELK; CHX tracking assay and ubiquitination assay detected the degradation of MELK on EZH2 ubiquitination. ChIP assay detected the enrichment of EZH2 and H3K27me3 on the LATS2 promoter region. LUAD cells were selected for in vitro validation, and the tumorigenic ability of LUAD cells was also observed in a transplantation tumour model of LUAD nude mice. MELK and EZH2 were highly expressed in LUAD samples, while LATS2 was lowly expressed. MELK interacted with EZH2 to inhibit its ubiquitination degradation; EZH2 elevated H3K27me3 modification in the LATS2 promoter to lower LATS2 expression. Silencing MELK or EZH2 or overexpressing LATS2 restrained LUAD cell proliferation and invasion, and facilitated their apoptosis. Silencing MELK or EZH2 or overexpressing LATS2 suppressed tumour formation in nude mice. This study demonstrated that MELK aggravated LUAD by upregulating EZH2 and downregulating LATS2.

Electrochemical reductive removal of trichloroacetic acids by a three-dimensional binderless carbon nanotubes/ CoP/Co foam electrode: Performance and mechanism.

Numerous chlorinated disinfection by-products (DBPs) are produced during the chlorination disinfection of water. Among them, chloroacetic acids (CAAs) are of great concern due to their potential human carcinogenicity. In this study, effective electrocatalytic dechlorination of trichloroacetic acids (TCAA), a typical CAAs, was achieved in the electrochemical system with the three-dimensional (3D) self-supported CoP on cobalt foam modified by carbon nanotubes (CNT/CoP/CF) as the cathode. At a 10 mA cm-2 current density, 74.5% of TCAA (500 µg L-1) was converted into AA within 100 min. In-situ growth of CoP increased the effective electrochemical surface area of the electrode. Electrodeposited CNT promoted electron transfer from the electrode surface to TCAA. Therefore, the production of surface-adsorbed atomic hydrogen (H*) on CNT/CoP/CF was improved, further resulting in excellent electrochemical dechlorination of TCAA. The dechlorination pathway of TCAA proceeded into acetic acids via direct electronic transfer and H*-mediated reduction on CNT/CoP/CF electrode. Additionally, the electroreduction efficiency of CNT/CoP/CF for TCAA exceeded 81.22% even after 20 cycles. The highly efficient TCAA reduction performance (96.57%) in actual water revealed the potential applicability of CNT/CoP/CF in the complex water matrix. This study demonstrated that the CNT/CoP/CF is a promising non-noble metal cathode to remove chlorinated DBPs in practice.

Evaluating the role of salinity in enhanced biogas production from two-stage anaerobic digestion of food waste by zero-valent iron.

Salts including NaCl are the most common food flavoring agents so they are often accumulated in food waste (FW) and have potential impact on anaerobic digestion (AD) of FW. In this study, the enhanced biogas production from two-stage anaerobic digestion (TSAD) of FW by microscale zero-valent iron (ZVI) under different salinity (3, 6, 9, and 15 g NaCl/L) was evaluated. Under salinity stress, ZVI becomes a continue-release electron donor due to the enhanced corrosion and dissolution effect and the slow-down surface passivation, further improving the performance of TSAD. Experimental results revealed that the biogas production including H2 and CH4 from TSAD with 10 g/L ZVI addition was promoted under salinity stress. The maximum H2 and CH4 yield (303.38 mL H2/g-VS and 253.84 mL CH4/g-VS) were observed at the salinity 9 g NaCl/L. Compared with that of zero salinity, they increased by 40.94% and 318.46%, respectively. Additionally, Sedimentibacter, an exoelectrogen that can participate in the direct interspecies electron transfer, also exhibited the highest relative abundance (34.96%) at the salinity 9 g NaCl/L. These findings obtained in this study might be of great importance for understanding the influence of salinity on the enhanced AD by ZVI.

Significance of circulating tumor cells detection in tumor diagnosis and monitoring.

To detect circulating tumor cells (CTCs) in the peripheral blood of patients with tumor, and to analyze the significance of CTC detection in tumor diagnosis and monitoring. In the present study, peripheral blood was collected from 125 patients with tumor, and CTCs were isolated and identified. Differences in CTC number and subtype detection were analyzed for different tumor diseases and stages. CTCs were detected in 122 of the 125 patients with tumor, with a positive rate of 97.6%. The number of CTCs increases in patients with vascular metastasis. The number of mesenchymal CTCs increases in patients with lymph node or vascular metastasis. The average ratio of epithelial CTCs in each positive sample decreases in the later stages of cancer compared with the earlier stages, while the average ratio of mesenchymal CTCs increases in the later stages of cancer compared with the earlier stages. The results showed that CTCs with mesenchymal phenotypes are closely related to lymph node or vascular metastasis. CTC detection can help with early diagnosis of tumor diseases. Continuous monitoring of changes in CTCs number and subtypes can assist clinical judgment of tumor disease development status and prognosis.

Drug resistance related genes in lung adenocarcinoma predict patient prognosis and influence the tumor microenvironment.

Lung adenocarcinoma (LUAD) is the predominant type of non-small lung cancer (NSCLC) with strong invasive ability and poor prognosis. The drug resistance related genes are potentially associated with prognosis of LUAD. Our research aimed to identify the drug resistance related genes and explore their potential prognostic value in LUAD patients. The data used in this study were obtained from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. Firstly, we screened drug resistance related genes in LUAD by differential gene analysis, univariate Cox regression and drug sensitivity analyses. Subsequently, we constructed a risk score model using LASSO Cox regression analysis, and verified whether the risk score can predict the survival of LUAD patients independent of other factors. Moreover, we explored the immune infiltration of 22 immune cells between high-risk and low-risk patients. Totally 10 drug-resistance positively related genes (PLEK2, TFAP2A, KIF20A, S100P, GDF15, HSPB8, SASH1, WASF3, LAMA3 and TCN1) were identified in LUAD. The risk score model of LUAD constructed with these 10 genes could reliably predict the prognosis of LUAD patients. 18 pathways were significantly activated in high-risk group compared with low-risk group. In addition, the infiltration proportion of multiple immune cells was significantly different between high-risk and low-risk groups, and the proportion of M1 phagocytes was significantly higher in the high-risk group compared with the low-risk group. The drug resistance related genes (PLEK2, TFAP2A, KIF20A, S100P, GDF15, HSPB8, SASH1, WASF3, LAMA3 and TCN1) could predict the prognosis of LUAD patients. Clarifying the roles and mechanisms of these 10 genes in regulating drug resistance in LUAD will help to improve individualized clinical treatment protocols and predict patient sensitivity to treatment.

Electroporation-Assisted Surface-Enhanced Raman Detection for Long-Term, Label-Free, and Noninvasive Molecular Profiling of Live Single Cells.

Molecule characterization of live single cells is greatly important in disease diagnoses and personalized treatments. Conventional molecule detection methods, such as mass spectrography, gene sequencing, or immunofluorescence, are usually destructive or labeled and unable to monitor the dynamic change of live cellular molecules. Herein, we propose an electroporation-assisted surface-enhanced Raman scattering (EP-SERS) method using a microchip to implement label-free, noninvasive, and continuous detections of the molecules of live single cells. The microchip containing microelectrodes with nanostructured EP-SERS probes has a multifunction of cell positioning, electroporation, and SERS detection. The EP-SERS method capably detects both the intracellular and extracellular molecules of live single cells without losing cell viability so as to enable long-term monitoring of the molecular pathological process in situ. We detect the molecules of single cells for two breast cancer cell lines with different malignancies (MCF-7 and MDA-MB-231), one liver cancer cell line (Huh-7), and one normal cell line (293T) using the EP-SERS method and classify these cell types to achieve high accuracies of 91.4-98.3% using their SERS spectra. Furthermore, 24 h continuous monitoring of the heterogeneous molecular responses of different cancer cell lines under doxorubicin treatment is successfully implemented using the EP-SERS method. This work provides a long-term, label-free, and biocompatible approach to simultaneously detect intracellular and extracellular molecules of live single cells on a chip, which would facilitate research and applications of cancer diagnoses and personalized treatments.

Computer-Vision-Based Dielectrophoresis Mobility Tracking for Characterization of Single-Cell Biophysical Properties.

Fast and precise measurements of live single-cell biophysical properties is significant in disease diagnosis, cytopathologic analysis, etc. Existing methods still suffer from unsatisfied measurement accuracy and low efficiency. We propose a computer vision method to track cell dielectrophoretic movements on a microchip, enabling efficient and accurate measurement of biophysical parameters of live single cells, including cell radius, cytoplasm conductivity, and cell-specific membrane capacitance, and in situ extraction of cell texture features. We propose a prediction-iteration method to optimize the cell parameter measurement, achieving high accuracy (less than 0.79% error) and high efficiency (less than 30 s). We further propose a hierarchical classifier based on a support vector machine and implement cell classification using acquired cell physical parameters and texture features, achieving high classification accuracies for identifying cell lines from different tissues, tumor and normal cells, different tumor cells, different leukemia cells, and tumor cells with different malignancies. The method is label-free and biocompatible, allowing further live cell studies on a chip, e.g., cell therapy, cell differentiation, etc.

Achieving high-performance electrocatalytic reduction of nitrate by N-rich carbon-encapsulated Ni-Cu bimetallic nanoparticles supported nickel foam electrode.

The cathode with low-energy consumption and long-term stability is pivotal to achieve the conversion of nitrate (NO3-) to nitrogen (N2) by electrocatalytic denitrification. Herein, a binder-free electrode was synthesized by directly immobilizing N-doped graphitized carbon layer-encapsulated NiCu bimetallic nanoparticles on nickel foam (NF) (NiCu@N-C/NF) and served as the cathode for electrocatalytic NO3- reduction. Morphological characterization indicated that Ni and Cu nanoparticles were encapsulated by the N-doped graphitized carbon layer and well-dispersed on the surface of NF. Compared with monometallic composite cathode (Cu@N-C/NF and Ni@N-C/NF), NiCu@N-C/NF exhibited better NO3- removal performance (98.63 %) and lower energy consumption (0.007 kW·h mmol-1), which should be attributed to its strong adsorption ability to NO3- and excellent electron transfer property. Meanwhile, its electrocatalytic performance could be maintained in wide initial NO3- concentration (1.79-7.14 mM) and solution pH (3-11). With the assistance of electrochlorination, the N2 selectivity of electrochemical system was up to 99.89 % in the presence of 0.028 M Cl-. More importantly, NiCu@N-C/NF electrode displayed an ultra-high stability during ten recycling experiments. This study indicated that the binderless composite cathode NiCu@N-C/NF had great potential in electrocatalytic NO3- removal from wastewater.

[Enhancement Effects and Mechanisms of Microscale Zero Valent Iron on the Performance of Anaerobic Co-digestion of Waste Activated Sludge and Food Waste].

Focusing on low biogas yields in the anaerobic co-digestion of waste activated sludge and food waste, the enhancing effects and mechanisms of microscale zero valent iron (mZVI) on anaerobic co-digestion was investigated. The results indicated that the addition of mZVI enhanced the methanogenesis stage of co-digestion but had no significant effect on the solubilization, hydrolysis, and acidification stages. With a dosage of 10 g·L-1 mZVI, the cumulative methane yield (based on VS) within 15 days reached 238.68 mL·g-1, which was 20.05% higher than the control group. The mechanism analysis showed that mZVI promoted electron transport system (ETS) activity (based on INTF/TS), which increased to 21.50 mg·(g·h)-1 with 10 g·L-1 mZVI compared to 13.43 mg·(g·h)-1 in the control group. Furthermore, mZVI enhanced direct interspecies electron transfer (DIET) between specific bacteria and methanogens. Microbial community analysis demonstrated that the abundance of DIET-related microorganisms, such as Syntrophomonas, Methanosarcina, and Methanobacterium, was higher in presence of mZVI.

Enhanced anaerobic co-digestion of waste activated sludge and food waste by sulfidated microscale zerovalent iron: Insights in direct interspecies electron transfer mechanism.

The enhancement of zerovalent iron (ZVI) on anaerobic digestion (AD) has been proved, but there are still some problems that constrain the large-scale application of ZVI, such as the destruction of cell membrane and the inhibition of methanogenesis led by rapid H2 accumulation. Aiming at these problems, sulfidated microscale zerovalent iron (S-mZVI) was employed to evaluate its effect on anaerobic co-digestion (AcoD) of waste activated sludge (WAS) and food waste (FW). Experimental results showed that S-mZVI promoted the direct interspecies electron transfer (DIET) between specific bacteria and methanogens, resulting in higher methane yield. At S-mZVI 10 g/L, the cumulative methane yield and ETS activity reached 264.78 mL/g-VS and 24.62 mg INTF/(g-TS h), which was 1.33 and 1.83 times that of blank. Microbiological analysis demonstrated that the abundance of DIET-related microorganisms such as Syntrophomonas, Methanosarcina and Methanobacterium increased with the increasing dosage of S-mZVI.

The inhibitory effect of thiosulfinate on volatile fatty acid and hydrogen production from anaerobic co-fermentation of food waste and waste activated sludge.

Thiosulfinate, a nature antibiotic, existed in all parts of Allium thereby accumulating in kitchen waste vastly. However, few literatures were available related to its influence on volatile fatty acids (VFA) and hydrogen production when kitchen waste digestion technology was applied. This study aimed to explore the inhibitory effect and the relevant mechanism. Experimental results showed that the hydrogen accumulation decreased from 23.2 ± 0.8 to 8.2 ± 0.1 mL/g VSS (volatile suspended solid) and maximal total VFA yield decreased from 765.7 ± 21.2 to 376.4 ± 21.7 mg COD (chemical oxygen demand)/g VSS when the dosage of thiosulfinate increased from 0 to 12.5 µg/g VSS. The mechanism study indicated, compared with control group, that the butyric acid decreased from 59% to 20.1% of total VFA yield when reactor in present of 12.5 µg/g VSS thiosulfinate. Moreover, the relative activities of functional enzymes were inhibited 73.4% (butyryl-CoA) and 72.7% (NADH), respectively.

Roles of MYC-targeting long non-coding RNA MINCR in cell cycle regulation and apoptosis in non-small cell lung Cancer.

BACKGROUND: Non-small cell lung cancer (NSCLC) is one of the leading causes of cancer death in the world, and has a relatively low survival rate. Long non-coding RNAs (lncRNAs) have been demonstrated to modulate cancer progression through a variety of molecular mechanisms. We sought to investigate the role and potential mechanism of MYC-induced long non-coding RNA (MINCR) in NSCLC. METHODS: Expression levels of MINCR was first identified using The Cancer Genome Atlas (TCGA), further confirmed with specimens from 29 NSCLC patients and three cell lines using qRT-PCR. Overexpression and knockdown of MINCR were performed in NSCLC cell lines through MINCR overexpression vectors and synthesized siRNAs, respectively. The roles of MINCR in NSCLC cell lines, such as cell proliferation, cell cycle arrest, and apoptosis, were identified by MTT, flow cytometry, and Western blot. The modulation of MINCR-regulated genes, including c-Myc and its downstream effectors, as well as apoptosis-associated genes, was analyzed using Western blot. RESULTS: MINCR expression was increased in NSCLC patients from TCGA datasets, and was also significantly increased in our collected specimens from NSCLC patients and NSCLC cell lines. Knocking down of MINCR greatly inhibited the growth of NSCLC cell lines PC9 and A549. In addition, silencing of MINCR induced cell cycle arrest and apoptosis. Furthermore, silencing of MINCR reduced the expression levels of oncogene c-Myc and its downstream cyclin A, cyclin D, CD4, and CDK2, as well as apoptosis-associated Bcl-2, while significantly increased the expression levels of cleaved PARP-1. In the meantime, overexpression of MINCR remarkably enhanced cell proliferation of PC9 cells and activated c-Myc and its downstream effectors. CONCLUSION: MINCR exerted inhibitory effects on the cell cycle arrest and apoptosis of NSCLC cells by activating c-Myc and its downstream effectors, suggesting that this lncRNA could be used as a potential therapeutic target for the treatment of NSCLC.

Increased IL-27/IL-27R expression in association with the immunopathology of murine ocular toxoplasmosis.

Interleukin 27 (IL-27) is a member of the IL-6/IL-12 family, and IL-27 receptor (IL-27R) consists of WSX-1 (the IL-27Rα subunit) and the signal-transducing subunit gp130. Human and mouse mast cells (MCs) express the IL-27R. To explore the expressions of IL-27/IL-27R subunits (WSX-1 and gp130) during acute ocular toxoplasmosis (OT), we established mouse model by intraocular injection of 500 Toxoplasma gondii RH strain tachyzoites. Histopathological changes were analyzed, MCs were counted by toluidine blue staining, and tryptase+/IL-27+ MCs were examined by immunofluorescence double-staining in the eyes and cervical lymph nodes (CLNs) of T. gondii-infected mice. The mRNA expressions of IL-27p28, WSX-1, gp130, and tachyzoite specific surface antigen 1 (SAG1) in the eyes and CLNs of T. gondii-infected mice, and the expressions of WSX-1 and gp130 in the murine mastocytoma cell line P815 infected with T. gondii tachyzoites in vitro were examined by using quantitative real-time reverse transcription-polymerase chain reaction. Our results showed that, after T. gondii infection, severe histopathological changes, increased numbers of total MCs and degranulated MCs, elevated expressions of IL-27p28, WSX-1, and gp130 were found in the eyes and CLNs, and significant correlations between the levels of IL-27 and SAG1 existed in the eyes and CLNs of T. gondii-infected mice. In addition, increased levels of WSX-1 and gp130 were examined in T. gondii-infected P815 cells. Our data suggested that IL-27/IL-27R expression induced by T. gondii infection may regulate MC-mediated immune response during acute OT in mouse model.

The first case of endophthalmitis due to Rhinocladiella basitona in an immunocompetent patient.

Rhinocladiella, a genus of black yeast-like fungi, is related to many infections in humans, including not only mild cutaneous lesions but also fatal brain infections. However, endophthalmitis caused by Rhinocladiella has never been reported by far. Herein, we present the first case of endophthalmitis due to Rhinocladiella basitona. The diagnosis was based on histopathology, mycology, and molecular identification. A 53-year-old female was struck by a piece of wood in her right eye. The wound in the central cornea became an ulcer and was aggravated continuously. Hyphae were found in the corneal scraping smear. Then endophthalmitis occurred and could not be controlled by the combined intravitreal antibiotic injections and vitrectomy. Finally, penetrating keratoplasty combined with retinal reattachment surgery was performed. Topical and systemic antifungal agents were administered for more than 1 month. The patient was cured, with improved visual acuity and clear corneal graft.

[Effect of extract of Ginkgo biloba on doxorubicin-associated cardiotoxicity in patients with breast cancer].

OBJECTIVE: To study the effect of extract of Ginkgo biloba (Egb761) on doxorubicin-associated cardiotoxicity in patients with breast cancer (BC). METHODS: Sixty BC patients in stage IV were randomly assigned to two groups, the control group was treated with chemotherapy, using 4 cycles of PA protocol alone and the treated group with the same chemotherapy and Egb761. Changes in electrocardiogram (ECG), myocardial enzyme spectrum (MES) and ultrasono-cardiogram (USCG) before and after treatment were observed. RESULTS: After treatment, the incidence of abnormal ECG was lower in the treated group than in the control group (6.7% vs 30.0%); significant differences were found between the two groups in the parameters of MES (P< 0.05); USCG showed significant difference between the two groups in left ventricular diastolic diameter (LVDd), left ventricular systolic diameter (LVDs), ratio of early and late diastolic transmitral peak flow velocity (E/A) and fractional shortening (FS), while there was no significant difference in ejection fraction (EF). CONCLUSION: Egb761 is an ideal drug for preventing and reducing the acute doxorbincin-induced cardiotoxicity; it could also be helpful for alleviating the chronic cardiotoxicity.