Human Hepatocytes with Drug Metabolic Function Induced from Fibroblasts by Lineage Reprogramming

SummaryObtaining fully functional cell types is a major challenge for drug discovery and regenerative medicine. Currently, a fundamental solution to this key problem is still lacking. Here, we show that functional human induced hepatocytes (hiHeps) can be generated from fibroblasts by overexpressing the hepatic fate conversion factors HNF1A, HNF4A, and HNF6 along with the maturation factors ATF5, PROX1, and CEBPA. hiHeps express a spectrum of phase I and II drug-metabolizing enzymes and phase III drug transporters. Importantly, the metabolic activities of CYP3A4, CYP1A2, CYP2B6, CYP2C9, and CYP2C19 are comparable between hiHeps and freshly isolated primary human hepatocytes. Transplanted hiHeps repopulate up to 30% of the livers of Tet-uPA/Rag2−/−/γc−/− mice and secrete more than 300 μg/ml human ALBUMIN in vivo. Our data demonstrate that human hepatocytes with drug metabolic function can be generated by lineage reprogramming, thus providing a cell resource for pharmaceutical applications

Unexpected values of Qs in the unconsolidated sediments of the Mississippi embayment

We studied the attenuation of shear waves at three sites in the Mississippi embayment using data recorded in boreholes drilled to depths of up to 60 m. The source was a highly repeatable compressed-air-driven hammer. To estimate attenuation we used a spectral ratio technique for fixed depth and variable frequency. The best-fit line for each depth z gives a measure of the cumulative attenuation, indicated by α(z). Then we fit a straight line to α(z) for a range of values of z. The slope of this line gives an estimate of the average attenuation per distance and was used to determine an average Qs. For one of the sites (Newport, northeastern Arkansas), Qs ranges between 34 (1.5 m ≤ z ≤ 44.2 m) and 44 (1.5 m ≤ z ≤ 51.8 m). These values are significantly higher than the more typical value of about 10 determined for unconsolidated sediments by other authors. In addition, these high values correspond to sediments with low average shear-wave velocity (about 300 m/sec). In contrast, average Qs and velocity for sediments in Shelby Forest (near Memphis, Tennessee) are 22 and 348 m/sec (22.6 m ≤ z ≤ 60.1 m), respectively. Therefore, these results go against the conventional wisdom that low velocity implies low Q. For the third site (Marked Tree, northeastern Arkansas), average Qs and velocity are 18 and 251 m/sec (9.8 m ≤ z ≤ 33.6 m), respectively. This site is about 75 km from Newport, and the differences in attenuation appear related to differences in lithology

A Simple and High Quality Method for Isolation and Extraction of Total RNA of Pholiota adipose

Analyzing functional values of RNA using RNA-seq technology is a hot spot of researches. In order to study the medicinal value of Pholiota adipose from the transcriptome level, it is necessary to extract and isolate RNA samples of high purity and high quality P. adipose. In this study, liquid nitrogen grinding Trizol one-step method was used to extract the total RNA of P. adipose. Quality test and statistical comparative analysis were carried out for RNA extract of liquid nitrogen grinding treated and untreated P. adipose. The results showed that the concentration of RNA in the samples treated with liquid nitrogen was much higher than that of the samples without grinding treatment. The OD260/280 of both was about 2, indicating that the purity of RNA was very high. Besides, the ratio of fluorescence intensity of 25S and 18S subunit strips of three replicate samples was 1.8, 1.9, and 1.9, close to 2, indicating RNA integrity is good. RIN test results of Agilent 2100 were 9.1, 8.7, and 9.3, higher than the standard value 6.8, further proving the integrity. In sum, liquid nitrogen grinding Trizol one-step method is a very simple and efficient method for extracting high quality total RNA of P. adipose

Guideline for the extraction, isolation, purification, and structural characterization of polysaccharides from natural resources

Abstract Polysaccharide is widely distributed in natural resources, and currently attracts scientists' attention for their various bio‐functions including immunomodulatory activity, hypoglycemic activity, hypolipidemic activity, antitumor activity, promoting effects on gastrointestinal function, and so on. The purity and structure characteristics of different polysaccharides greatly restrict the in‐depth study of their bioactivity mechanism. Therefore, this guideline will display the extraction, isolation, purification, and structural characterization of polysaccharides from natural resources. The extraction methods of acid‐soluble polysaccharides, water‐soluble polysaccharides, and alkaline soluble polysaccharides are included. Common purification methods like membrane filtration, alcohol precipitation, and column chromatographic separation are explained in detail. A comprehensive procedure for polysaccharide structural characterization is given from molecular weight and monosaccharide composition study to Fourier transform infrared and nuclear magnetic resonance analysis. Examples are added where necessary to help you better understand this guideline

Dilatancy Characteristics and Constitutive Modelling of the Unsaturated Soil Based on Changes in the Mass Water Content

Most soil mechanics theories are limited to strain hardening and shrinkage under high compressive stresses, and there are some shortcomings in the selection of suction or degree of saturation as the water content state varies in the constitutive models of unsaturated soil. Based on the triaxial shear tests of unsaturated compacted soil (a silt of high plasticity) with different water content and confining pressure (low-confining), a shear dilatancy model of unsaturated soil based on the mass water content is proposed in this paper. The influence of the water content on the shear deformation characteristics of the unsaturated soil is analysed. The stress–dilatancy relationship and the prediction equation of the minimum dilatancy rate of the unsaturated soil under different water content and different confining pressure are provided. Selecting the mass water content as the state variable, a constitutive model suitable for the dilatancy of unsaturated soil is established. The method of determining model parameters based on the mass water content is analysed. The applicability of the model is verified by comparisons between the predicted and experimental results

Fabrication and properties of strip casting 4.5 wt% Si steel thin sheet

Three 4.5 wt% Si steel thin sheets with different thicknesses were efficiently fabricated by twin-roll strip casting, warm rolling and cold rolling followed by final annealing. A comprehensive investigation from the workability of the as-cast strip to the magnetic property of the produces was performed to illustrate the superiority of the new materials. The results show that the as-cast strip, which has a much lower Vickers hardness than that of the 6.5 wt% Si steel, is suitable for rolling processing. The X-ray diffraction (XRD) and transmission electron microscopy (TEM) studies confirm that no ordering phase exists in the as-cast strip. The cold-rolled thin sheets exhibit good surface quality without edge cracks. Furthermore, all the three 4.5 wt% Si steel thin sheets possess relative strong //ND texture and present high magnetic inductions and low iron losses after finial annealing

Boosting the Oxygen Evolution Reaction by Controllably Constructing FeNi₃/C Nanorods

Transition bimetallic alloy-based catalysts are regarded as attractive alternatives for the oxygen evolution reaction (OER), attributed to their competitive economics, high conductivity and intrinsic properties. Herein, we prepared FeNi3/C nanorods with largely improved catalytic OER activity by combining hydrothermal reaction and thermal annealing treatment. The temperature effect on the crystal structure and chemical composition of the FeNi3/C nanorods was revealed, and the enhanced catalytic performance of FeNi3/C with an annealing temperature of 400 °C was confirmed by several electrochemical tests. The outstanding catalytic performance was assigned to the formation of bimetallic alloys/carbon composites. The FeNi3/C nanorods showed an overpotential of 250 mV to afford a current density of 10 mA cm−2 and a Tafel slope of 84.9 mV dec−1, which were both smaller than the other control samples and commercial IrO2 catalysts. The fast kinetics and high catalytic stability were also verified by electrochemical impendence spectroscopy and chronoamperometry for 15 h. This study is favorable for the design and construction of bimetallic alloy-based materials as efficient catalysts for the OER

Use of Ferritin Expression, Regulated by Neural Cell-Specific Promoters in Human Adipose Tissue-Derived Mesenchymal Stem Cells, to Monitor Differentiation with Magnetic Resonance Imaging In Vitro.

The purpose of this study was to establish a method for monitoring the neural differentiation of stem cells using ferritin transgene expression, under the control of a neural-differentiation-inducible promoter, and magnetic resonance imaging (MRI). Human adipose tissue-derived mesenchymal stem cells (hADMSCs) were transduced with a lentivirus containing the human ferritin heavy chain 1 (FTH1) gene coupled to one of three neural cell-specific promoters: human synapsin 1 promoter (SYN1p, for neurons), human glial fibrillary acidic protein promoter (GFAPp, for astrocytes), and human myelin basic protein promoter (MBPp, for oligodendrocytes). Three groups of neural-differentiation-inducible ferritin-expressing (NDIFE) hADMSCs were established: SYN1p-FTH1, GFAPp-FTH1, and MBPp-FTH1. The proliferation rate of the NDIFE hADMSCs was evaluated using a Cell Counting Kit-8 assay. Ferritin expression was assessed with western blotting and immunofluorescent staining before and after the induction of differentiation in NDIFE hADMSCs. The intracellular iron content was measured with Prussian blue iron staining and inductively coupled plasma mass spectrometry. R2 relaxation rates were measured with MRI in vitro. The proliferation rates of control and NDIFE hADMSCs did not differ significantly (P > 0.05). SYN1p-FTH1, GFAPp-FTH1, and MBPp-FTH1 hADMSCs expressed specific markers of neurons, astrocytes, and oligodendrocytes, respectively, after neural differentiation. Neural differentiation increased ferritin expression twofold, the intracellular iron content threefold, and the R2 relaxation rate two- to threefold in NDIFE hADMSCs, resulting in notable hypointensity in T2-weighted images (P < 0.05). These results were cross-validated. Thus, a link between neural differentiation and MRI signals (R2 relaxation rate) was established in hADMSCs. The use of MRI and neural-differentiation-inducible ferritin expression is a viable method for monitoring the neural differentiation of hADMSCs

EVLncRNAs: a manually curated database for long non-coding RNAs validated by low-throughput experiments

Long non-coding RNAs (lncRNAs) play important functional roles in various biological processes. Early databases were utilized to deposit all lncRNA candidates produced by high-throughput experimental and/or computational techniques to facilitate classification, assessment and validation. As more lncRNAs are validated by low-throughput experiments, several databases were established for experimentally validated lncRNAs. However, these databases are small in scale (with a few hundreds of lncRNAs only) and specific in their focuses (plants, diseases or interactions). Thus, it is highly desirable to have a comprehensive dataset for experimentally validated lncRNAs as a central repository for all of their structures, functions and phenotypes. Here, we established EVLncRNAs by curating lncRNAs validated by low-throughput experiments (up to 1 May 2016) and integrating specific databases (lncRNAdb, LncRANDisease, Lnc2Cancer and PLNIncRBase) with additional functional and disease-specific information not covered previously. The current version of EVLncRNAs contains 1543 lncRNAs from 77 species that is 2.9 times larger than the current largest database for experimentally validated lncRNAs. Seventy-four percent lncRNA entries are partially or completely new, comparing to all existing experimentally validated databases. The established database allows users to browse, search and download as well as to submit experimentally validated lncRNAs. The database is available at here