Intracellular Trafficking Network of Protein Nanocapsules: Endocytosis, Exocytosis and Autophagy

The inner membrane vesicle system is a complex transport system that includes endocytosis, exocytosis and autophagy. However, the details of the intracellular trafficking pathway of nanoparticles in cells have been poorly investigated. Here, we investigate in detail the intracellular trafficking pathway of protein nanocapsules using more than 30 Rab proteins as markers of multiple trafficking vesicles in endocytosis, exocytosis and autophagy. We observed that FITC-labeled protein nanoparticles were internalized by the cells mainly through Arf6-dependent endocytosis and Rab34-mediated micropinocytosis. In addition to this classic pathway: early endosome (EEs)/late endosome (LEs) to lysosome, we identified two novel transport pathways: micropinocytosis (Rab34 positive)-LEs (Rab7 positive)-lysosome pathway and EEs-liposome (Rab18 positive)-lysosome pathway. Moreover, the cells use slow endocytosis recycling pathway (Rab11 and Rab35 positive vesicles) and GLUT4 exocytosis vesicles (Rab8 and Rab10 positive) transport the protein nanocapsules out of the cells. In addition, protein nanoparticles are observed in autophagosomes, which receive protein nanocapsules through multiple endocytosis vesicles. Using autophagy inhibitor to block these transport pathways could prevent the degradation of nanoparticles through lysosomes. Using Rab proteins as vesicle markers to investigation the detail intracellular trafficking of the protein nanocapsules, will provide new targets to interfere the cellular behaver of the nanoparticles, and improve the therapeutic effect of nanomedicine

eIF4A inhibitors suppress cell-cycle feedback response and acquired resistance to CDK4/6 inhibition in cancer

CDK4/6 inhibitors are FDA-approved drugs for estrogen receptor-positive (ER+) breast cancer and are being evaluated to treat other tumor types, including KRAS-mutant non-small cell lung cancer (NSCLC). However, their clinical utility is often limited by drug resistance. Here, we sought to better understand the resistant mechanisms and help devise potential strategies to overcome this challenge. We show that treatment with CDK4/6 inhibitors in both ER+ breast cancer and KRAS-mutant NSCLC cells induces feedback upregulation of cyclin D1, CDK4, and cyclin E1, mediating drug resistance. We demonstrate that rocaglates, which preferentially target translation of key cell-cycle regulators, effectively suppress this feedback upregulation induced by CDK4/6 inhibition. Consequently, combination treatment of CDK4/6 inhibitor palbociclib with the eukaryotic initiation factor (eIF) 4A inhibitor, CR-1-31-B, is synergistic in suppressing the growth of these cancer cells in vitro and in vivo Furthermore, ER+ breast cancer and KRAS-mutant NSCLC cells that acquired resistance to palbociclib after chronic drug exposure are also highly sensitive to this combination treatment strategy. Our findings reveal a novel strategy using eIF4A inhibitors to suppress cell-cycle feedback response and to overcome resistance to CDK4/6 inhibition in cancer.Accepted manuscrip

Total saponins from Trillium tschonoskii Maxim promote neurological recovery in model rats with post-stroke cognitive impairment

Total saponins from Trillium tschonoskii Maxim (TSTT), a bioactive component of local natural herbs in the Enshi area, China, have been demonstrated to have functions of restoring cognitive capacity and promoting axonal regeneration post-stroke, but the mechanism of this process remains unclear. The hippocampus is a critical tissue for controlling learning and memory capacity, and the sonic hedgehog (Shh) signaling pathway plays a major role in the patterning and synaptic plasticity of hippocampal neural circuits. Therefore, we aimed to investigate whether TSTT could restore learning and cognitive functions by modulating the Shh pathway in rats with post-stroke cognitive impairment (PSCI). The ischemia model was established by permanent middle cerebral artery occlusion (MCAO) in 100 Sprague–Dawley (SD) rats, and the model rats were administered using TSTT (100 mg/kg) or donepezil hydrochloride as the positive control (daily 0.45 mg/kg, DON) for 4 weeks after the operation. As assessed by the Morris water maze test, the cognitive function of PSCI rats was significantly improved upon TSTT treatment. Meanwhile, the cerebral infarct volume reduced with TSTT, as shown by HE and TTC staining, and the number of Nissl bodies and dendritic spine density were significantly increased, as shown by Nissl and Golgi staining. In addition, TSTT upregulated PSD-95, SYN, and GAP-43, and inhibited neuronal apoptosis, as evidenced by increased Bcl-2 levels along with decreased Bax and caspase-3 expression. TSTT could also significantly upregulate Shh, Ptch1, Smo, and Gli1 proteins, indicating the activation of the Shh signaling pathway. Therefore, TSTT can protect PSCI rats by inhibiting apoptosis and promoting neuronal synaptic remodeling. The Shh pathway is also involved

SMARCB1 loss induces druggable cyclin D1 deficiency via upregulation of MIR17HG in atypical teratoid rhabdoid tumors

Atypical teratoid rhabdoid tumor (ATRT) is a fatal pediatric malignancy of the central neural system lacking effective treatment options. It belongs to the rhabdoid tumor family and is usually caused by biallelic inactivation of SMARCB1, encoding a key subunit of SWI/SNF chromatin remodeling complexes. Previous studies proposed that SMARCB1 loss drives rhabdoid tumor by promoting cell cycle through activating transcription of cyclin D1 while suppressing p16. However, low cyclin D1 protein expression is observed in most ATRT patient tumors. The underlying mechanism and therapeutic implication of this molecular trait remain unknown. Here, we show that SMARCB1 loss in ATRT leads to the reduction of cyclin D1 expression by upregulating MIR17HG, a microRNA (miRNA) cluster known to generate multiple miRNAs targeting CCND1. Furthermore, we find that this cyclin D1 deficiency in ATRT results in marked in vitro and in vivo sensitivity to the CDK4/6 inhibitor palbociclib as a single agent. Our study identifies a novel genetic interaction between SMARCB1 and MIR17HG in regulating cyclin D1 in ATRT and suggests a rationale to treat ATRT patients with FDA- approved CDK4/6 inhibitors. © 2020 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/156416/2/path5493.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/156416/1/path5493_am.pd

Improved IMPES Scheme for the Simulation of Incompressible Three-Phase Flows in Subsurface Porous Media

In this work, an improved IMplicit Pressure and Explicit Saturation (IMPES) scheme is proposed to solve the coupled partial differential equations to simulate the three-phase flows in subsurface porous media. This scheme is the first IMPES algorithm for the three-phase flow problem that is locally mass conservative for all phases. The key technique of this novel scheme relies on a new formulation of the discrete pressure equation. Different from the conventional scheme, the discrete pressure equation in this work is obtained by adding together the discrete conservation equations of all phases, thus ensuring the consistency of the pressure equation with the three saturation equations at the discrete level. This consistency is important, but unfortunately it is not satisfied in the conventional IMPES schemes. In this paper, we address and fix an undesired and well-known consequence of this inconsistency in the conventional IMPES in that the computed saturations are conservative only for two phases in three-phase flows, but not for all three phases. Compared with the standard IMPES scheme, the improved IMPES scheme has the following advantages: firstly, the mass conservation of all the phases is preserved both locally and globally; secondly, it is unbiased toward all phases, i.e., no reference phases need to be chosen; thirdly, the upwind scheme is applied to the saturation of all phases instead of only the referenced phases; fourthly, numerical stability is greatly improved because of phase-wise conservation and unbiased treatment. Numerical experiments are also carried out to demonstrate the strength of the improved IMPES scheme

pH-shift strategy improving the thermal stability and oxidation stability of rice starch/casein-based high internal phase emulsions for the application in fish cake

The thermal stability of the different pH-shift rice starch/casein-based high internal phase emulsions (SC-HIPE) were evaluated in the present study to verify potential in improving the quality of fish cake. The results showed that the pH-shift treatment improved thermal stability (from 27.23% to 76.33%) and oxidation time (from 5.01 h to 6.86 h) of SC-HIPE, which showed the smaller droplet size (decreased from 15.14 to 1.64 μm) and higher storage module. The breaking force of FC with thermal stable SC-HIPE (average 64.95 g) was higher than that with thermal unstable SC-HIPE (51.05 g). The cohesiveness, adhesiveness and chewiness could be improved by adding thermal stable SC-HIPE, compared with pork fat. Additionally, combining sensory evaluation, the thermal stable SC-HIPE improved the gel quality, thus it could be completely replaced pork fat in the preparation of FC, which provided theoretical guidance for the preparation and application of fat substitutes

Satellite Positioning and Orbit Determination System SPODS:Theory and Test

The Satellite Positioning and Orbit Determination System(SPODS)is a software package for GNSS positioning/orbit determination,developed by the Xi'an Research Institute of Surveying and Mapping.So far it has been able to treat GPS data and has the capability of high precision GPS positioning and orbit determination.The underlying theory and the performance test are briefly addressed.The test utilizes the GPS data collected from some 127IGS stations during days 4~10of 2009.The results show that the rms 1D difference is 1.1cm between SPODS orbits and final IGS combined orbits,and that the repeatability of daily solutions of station coordinates is 1.5mm for horizontal components,and 4.5mm for height component,and that the consistency of ERP solutions with final IGS values is 0.025mas,0.093mas and 0.013ms/d respectively for pole coordinates and changes in length of day