Epithelium-Specific ETS (ESE)-1 upregulated GP73 expression in hepatocellular carcinoma cells

Background: Golgi protein-73 (GP73) is a Golgi transmembrane glycoprotein elevated in numerous liver diseases. Clinically, GP73 is strongly elevated in the serum of HCC patients and is thus regarded as a novel potential biomarker for HCC. However, the mechanism leading to GP73 dysregulation in liver diseases remains unknown. Results: This study determined that epithelium-specific ETS (ESE)-1, an epithelium-specific transcription factor, and GP73 expressions were induced by IL-1β stimulation in vitro, and both were triggered during liver inflammation in vivo. In hepatocellular carcinoma cells, the overexpression of ESE-1 induced GP73 expression, whereas its knock-down did the opposite. Mechanistically, ESE-1 activated GP73 expression by directly binding to its promoter. Conclusions: Our findings supported a novel paradigm for ESE-1 as a transcriptional mediator of GP73. This study provided a possible mechanism for GP73 upregulation in liver diseases. Electronic supplementary material The online version of this article (doi:10.1186/2045-3701-4-76) contains supplementary material, which is available to authorized users

The Golgi Localization of GOLPH2 (GP73/GOLM1) Is Determined by the Transmembrane and Cytoplamic Sequences

Golgi phosphoprotein 2 (GOLPH2) is a resident Golgi type-II membrane protein upregulated in liver disease. Given that GOLPH2 traffics through endosomes and can be secreted into the circulation, it is a promising serum marker for liver diseases. The structure of GOLPH2 and the functions of its different protein domains are not known. In the current study, we investigated the structural determinants for Golgi localization using a panel of GOLPH2 truncation mutants. The Golgi localization of GOLPH2 was not affected by the deletion of the C-terminal part of the protein. A truncated mutant containing the N-terminal portion (the cytoplasmic tail and transmembrane domain (TMD)) localized to the Golgi. Sequential deletion analysis of the N-terminal indicated that the TMD with a positively charged residue in the cytoplasmic N-terminal tail were sufficient to support Golgi localization. We also showed that both endogenous and secreted GOLPH2 exist as a disulfide-bonded dimer, and the coiled-coil domain was sufficient for dimerization. This structural knowledge is important for the understanding the pathogenic role of GOLPH2 in liver diseases, and the development of GOLPH2-based hepatocellular cancer diagnostic methods

An Algorithm and Implementation Based on an Agricultural EOQ Model

With the improvement of living quality, the agricultural supermarket gradually take the place of the farmers market as the trend. But the agricultural supermarkets’ inappropriate inventory strategies are wasteful and inefficient. So this paper will put forward an inventory strategy for the agricultural supermarkets to lead the conductor decides when and how much to shelve the product. This strategy has significant meaning that it can reduce the loss and get more profit. The research methods are based on the inventory theory and the EOQ model, but the authors add multiple cycles’ theory to them because of the agricultural products’ decreasing characteristics. The research procedures are shown as follows. First, the authors do research in the agricultural supermarket to find their real conduction, and then put forward the new strategy in this paper. Second, the authors found out the model. At last, the authors search the specialty agriculture document to find the data such as the loss rate and the fresh parameters, and solve it out by MATLAB. The numerical result proves that the strategy is better than the real conduction in agricultural supermarket, and it also proves the feasibility

An Algorithm and Implementation Based on an Agricultural EOQ Model

With the improvement of living quality, the agricultural supermarket gradually take the place of the farmers market as the trend. But the agricultural supermarkets’ inappropriate inventory strategies are wasteful and inefficient. So this paper will put forward an inventory strategy for the agricultural supermarkets to lead the conductor decides when and how much to shelve the product. This strategy has significant meaning that it can reduce the loss and get more profit. The research methods are based on the inventory theory and the EOQ model, but the authors add multiple cycles’ theory to them because of the agricultural products’ decreasing characteristics. The research procedures are shown as follows. First, the authors do research in the agricultural supermarket to find their real conduction, and then put forward the new strategy in this paper. Second, the authors found out the model. At last, the authors search the specialty agriculture document to find the data such as the loss rate and the fresh parameters, and solve it out by MATLAB. The numerical result proves that the strategy is better than the real conduction in agricultural supermarket, and it also proves the feasibility

GP73 is upregulated by hepatitis C virus (HCV) infection and enhances HCV secretion.

Hepatitis C virus (HCV) is a major cause of chronic liver disease. However, little is known about the details of its assembly and secretion. Golgi-related proteins have been recently proven to have a key function in HCV secretion. Golgi protein 73 (GP73), a resident Golgi membrane protein, is a potential serum biomarker for the diagnosis of liver diseases and hepatocellular carcinoma. Previous studies have demonstrated the upregulation of GP73 in the liver samples and sera of HCV-infected patients. However, the function and regulatory mechanism of GP73 in HCV infection at the cellular level remain unknown. In this study, we examined the expression level of GP73 in HCV infected cells and its effect on HCV life cycle in cell culture systems. Both the protein expression and mRNA levels of GP73 significantly increased in HCV subgenomic replicon-harboring cells and HCV-infected cells, which imply that GP73 was upregulated by HCV infection. HCV production was significantly enhanced when GP73 was overexpressed, but dramatically inhibited when GP73 was silenced. However, the overexpression and knockdown of GP73 showed no evident effect on the entry, protein translation, RNA replication, and assembly of HCV, which indicates that GP73 enhanced the secretion process. Moreover, the coiled-coil domain of GP73 was required to increase HCV secretion. GP73 increased and interacted with apolipoprotein E, an identified host factor that assists in HCV secretion. These results demonstrate the critical function of GP73 in HCV secretion and provide new insights into the therapeutic design of antiviral strategies

Preparation of supported carbon molecular sieve membrane from novolac phenol-formaldehyde resin

An asymmetric carbon membrane was prepared by coating alcohol solution of novolac phenol-formaldehyde resin containing a little hexamine on a porous resin support from the same material. After drying in air for two days at room temperature, the coated support was heated at 150 degrees C for I h in air (heating rate: 0.5 degrees C/min) and then carbonized at 800 degrees C (heating rate: 0.5 degrees C/min) in Ar atmosphere. The support and the membrane layer were carbonized simultaneously. The coating-pyrolysis cycle only needed one time. SEM photographs showed the carbon membrane had an asymmetric structure formed by a dense skin layer with a thickness of around 35 mu m and a porous substrate. Pure gases of different molecular size (H-2, CO2, O-2, N-2 and CH4) were used to test the carbon membrane permeance property. The membrane has a good selectivity for H-2/N-2 and H-2/CH4 with H-2 permeance of 4.05 x 10(-6) cm(3) cm(-2) s(-1) cmHg(-1). The permeance is independent of pressure. The results indicate that the gases transport through the membrane according to molecular sieve mechanism. (c) 2007 Elsevier B.V. All rights reserved

Provably Efficient Iterated CVaR Reinforcement Learning with Function Approximation

Risk-sensitive reinforcement learning (RL) aims to optimize policies that balance the expected reward and risk. In this paper, we investigate a novel risk-sensitive RL formulation with an Iterated Conditional Value-at-Risk (CVaR) objective under linear and general function approximations. This new formulation, named ICVaR-RL with function approximation, provides a principled way to guarantee safety at each decision step. For ICVaR-RL with linear function approximation, we propose a computationally efficient algorithm ICVaR-L, which achieves an $\widetilde{O}(\sqrt{\alpha^{-(H+1)}(d^2H^4+dH^6)K})$ regret, where $\alpha$ is the risk level, $d$ is the dimension of state-action features, $H$ is the length of each episode, and $K$ is the number of episodes. We also establish a matching lower bound $\Omega(\sqrt{\alpha^{-(H-1)}d^2K})$ to validate the optimality of ICVaR-L with respect to $d$ and $K$. For ICVaR-RL with general function approximation, we propose algorithm ICVaR-G, which achieves an $\widetilde{O}(\sqrt{\alpha^{-(H+1)}DH^4K})$ regret, where $D$ is a dimensional parameter that depends on the eluder dimension and covering number. Furthermore, our analysis provides several novel techniques for risk-sensitive RL, including an efficient approximation of the CVaR operator, a new ridge regression with CVaR-adapted features, and a refined elliptical potential lemma.Comment: 27 pages, 3 figure

GP73 enhances HCV production.

<p>(<b>A</b>)The mRNA levels of GP73 in stable cell lines were detected by qRT-PCR. (<b>B</b>) Infection efficiency of HCV-GFP in Huh7.5.1 stable cells was detected with flow cytometry at the indicated time points after infection at 0.02 MOI. (<b>C</b>) Intracellular viral RNA in Huh7.5.1 cells was quantified by qRT-PCR. The values are normalized to those of 24 h post-infection. (<b>D)</b> HCV-GFP infectivity of the supernatant at 72 h and 96 h post-infection were assayed by flow cytometry. (<b>E</b>) HCVpp infectivity in stable cells. The values are represented relative to naive Huh7.5.1 after normalization with VSVGpp infectivity. (<b>F</b>) HCV RNA levels in stable cells at 96 h post-transfection with pSGR-JFH1. The values are represented relative to those of naive Huh7.5.1 after normalization with HCV RNA level at 6 h post-transfection. The results are presented as mean ± SEM derived from three experiments (*<i>P</i><0.05; **<i>P</i><0.01).</p

GP73 increases the secretion of HCV through the coiled-coil domain.

<p>Huh7.5.1 cells infected with HCV-GFP at 0.02 MOI were transfected with GP73 truncation expression plasmids at 48 h post-infection. At 48 h after transfection, Huh7.5.1 cells were harvested and assayed. (<b>A</b>) Protein expression levels of GP73 truncations were detected by western blot. Infection efficiency (<b>B</b>) and MFI (<b>C</b>) of HCV-infected Huh7.5.1 were assayed by flow cytometry (<b>D</b>) Supernatant HCV-GFP titer was assayed by flow cytometry. (<b>E</b>) HCV viral RNA in the supernatant was quantified by qRT-PCR. (<b>F</b>) HCV-GFP titer of the supernatant from Huh7.5.1 cells transfected with indicated plasmids was assayed by flow cytometry. (<b>G</b>) Intracellular HCV-GFP infectivity was assayed by flow cytometry. (<b>H</b>) Intracellular HCV RNA levels were quantified by qRT-PCR. (<b>I</b>) Diagram of GP73 truncation structure and summary of effects on HCV production. (Y: Yes; N: No; UP: Upregulated). The results are presented as mean ± SEM derived from three experiments (*<i>P</i><0.05; **<i>P</i><0.01).</p

GP73 increases HCV production in the supernatant.

<p>(<b>A</b>) Schematic of the experimental procedure. Huh7.5.1 cells were infected with the lentivirus carrying the coding sequence of GP73 (pRlenti-GP73), vector control (pRlenti), shRNA expression cassette expressing shRNA against GP73 (shGP73-1 and shGP73-2), or non-target control (shNT) for 6 h. After 24 h, Huh7.5.1 cells were infected with HCV-GFP at 0.02 MOI for another 6 h. The GP73 mRNA levels (<b>B</b>), GP73 protein levels (<b>C</b>), the infection efficiency (<b>D</b>), and MFI (<b>E</b>) of Huh7.5.1 cells were assayed 72 h after infection with HCV-GFP. The HCV-GFP titer (<b>F</b>) and HCV viral RNA levels (<b>G</b>) in the supernatant were assayed by flow cytometry or qRT-PCR. The value of pRlenti-GP73 is represented relative to that of pRlenti. The values of shRNAs against GP73 are represented relative to those of shNT. The results are presented as mean ± SEM derived from three experiments (*P<0.05; **P<0.01).</p