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Background Pradefovir is a liver-targeted prodrug of adefovir, a nucleoside/nucleotide analogue with antiviral activity against hepatitis B virus (HBV) DNA polymerase. This phase 2 study compared the efficacy and safety of oral pradefovir (30, 45, 60, or 75 mg) versus tenofovir disoproxil fumarate (TDF; 300 mg) and aimed to identify the most appropriate dose of pradefovir for the forthcoming phase 3 study. Methods Treatment-naive and experienced (not on treatment >6 months) patients with chronic hepatitis B were eligible. Results A total of 240 participants were randomized and treated in the study (48 per group). Approximately 80% were hepatitis B e antigen (HBeAg) positive, and 10% had liver cirrhosis. The reductions from baseline in HBV DNA levels achieved at week 24 were 5.40, 5.34, 5.33, and 5.40 log10 IU/mL, with pradefovir doses of 30-, 45-, 60-, and 75-mg, respectively, compared with 5.12 log10 IU/mL with TDF. However, HBeAg loss was attained by more participants who received 45-, 60-, or 75-mg pradefovir than by those receiving TDF (12%, 6%, and 9% vs 3%). The TDF group exhibited a more significant increase in serum creatinine than the pradefovir 30- and 45-mg groups, and serum phosphate levels were comparable among all groups. Most adverse events (AEs) were mild (grade 1). No treatment-related severe AEs were reported. Overall, AEs and laboratory abnormalities were comparable to those in the TDF group. Conclusions Pradefovir and TDF exhibited comparable reductions in HBV DNA levels. All treatments were safe and well tolerated. Clinical Trials registration NCT00230503 and China Drug Trials CTR2018042

Exploring the expression of miR-571 in patients with liver fibrosis and its role in the progression of liver fibrosis. A total of 74 patients with liver fibrosis in our institution from September to December 2018 were collected for study, and the expression of miR-571, Notch3 and Jagged1 in patients with different progressions of liver fibrosis was determined by RT-PCR and Western blot analysis. Set up Notch3 up group and Notch3 down regulated group, RT-PCR and Western blot were used to determine the effect of Notch signaling on the expression of fibrogenic factors. CCK-8, cell scratch assays, Transwell assays, flow cytometry were used to determine the effect of miR-571 on LX-2 proliferation, migration, apoptosis in human stem stellate cells, and RT-PCR, Western blot assays were performed to determine the effect of miR-571 on the Notch3 signaling pathway and the expression of profibrogenic factors. miR-571, Notch3 and Jagged1 are up-regulated in patients with liver fibrosis and is associated with the progression of liver fibrosis. Notch3 signaling pathway can promote the expression of fibroblast in human hepatic stellate cells; miR-571 can inhibit the apoptosis of human hepatic stellate cells, promote cell proliferation and migration; up regulation of miR-571 can promote the expression of Notch3 and Jagged1, and up-regulation of miR-571 also promoted the expression of related fibroblasts. MiR-571 can promote the activation of human stem cell stellate cells and the expression of fibroblast related factors through Notch3 signaling pathway.

Aiming to make full use of three kinds of solid wastes from calcium carbide industry, this paper first investigated elements and mineral compounds of them through inductively coupled plasma and X-ray diffraction (XRD). The three solid waste samples, removed dust (RD), coke dust (CD) and purified dust (PD), have higher Ca/S molar ratios than conventional fuels such as coal. In particular, Ca/S molar ratio of PD unexpectedly achieved 28.05. Calcium in PD is present mainly in the form of calcium oxides, but little calcium carbonates, which was contrary to calcium in RD. In addition, the large proportion of fixed carbon in the samples CD and RD was amorphous carbon. Besides, three former dust samples were mixed at a specific ratio to obtain a new sample, mixed dust (MD), for further experiments as a reference of future industrial applications. Combining thermogravimetric system and Fourier transform infrared spectroscopy system helped to investigate the combustion process of all four samples in air atmosphere. Large amounts of reactive amorphous carbon identified by XRD desorb pollutants adsorbed in retorting process firstly and then help the samples RD, CD and MD ignite easily, which indicates they have potential to be fuels. Besides, the calcium oxide catalyzed the NOx formation during the combustion, which should be inhibited.

BACKGROUND Liver fibrosis is a refractory disease whose persistence can eventually induce cirrhosis or even liver cancer. Early liver fibrosis is reversible by intervention. As a member of the transforming growth factor-beta (TGF-β) superfamily, bone morphogenetic protein 7 (BMP7) has anti-liver fibrosis functions. However, little is known about BMP7 expression changes and its potential regulatory mechanism as well as the relationship between BMP7 and TGF-β during liver fibrosis. In addition, the mechanism underlying the anti-liver fibrosis function of BMP7 needs to be further explored. AIM To investigate changes in the dynamic expression of BMP7 during liver fibrosis, interactions between BMP7 and TGF-β1, and possible mechanisms underlying the anti-liver fibrosis function of BMP7. METHODS Changes in BMP7 expression during liver fibrosis and the interaction between BMP7 and TGF-β1 in mice were observed. Exogenous BMP7 was used to treat mouse primary hepatic stellate cells (HSCs) to observe its effect on activation, migration, and proliferation of HSCs and explore the possible mechanism underlying the anti-liver fibrosis function of BMP7. Mice with liver fibrosis received exogenous BMP7 intervention to observe improvement of liver fibrosis by using Masson’s trichrome staining and detecting the expression of the HSC activation indicator alpha-smooth muscle actin (α-SMA) and the collagen formation associated protein type I collagen (Col I). Changes in the dynamic expression of BMP7 during liver fibrosis in the human body were further observed. RESULTS In the process of liver fibrosis induced by carbon tetrachloride (CCl4) in mice, BMP7 protein expression first increased, followed by a decrease; there was a similar trend in the human body. This process was accompanied by a sustained increase in TGF-β1 protein expression. In vitro experiment results showed that TGF-β1 inhibited BMP7 expression in a time- and dose-dependent manner. In contrast, high doses of exogenous BMP7 inhibited TGF-β1-induced activation, migration, and proliferation of HSCs; this inhibitory effect was associated with upregulation of pSmad1/5/8 and downregulation of phosphorylation of Smad3 and p38 by BMP7. In vivo experiment results showed that exogenous BMP7 improved liver fibrosis in mice. CONCLUSION During liver fibrosis, BMP7 protein expression first increases and then decreases. This changing trend is associated with inhibition of BMP7 expression by sustained upregulation of TGF-β1 in a time- and dose-dependent manner. Exogenous BMP7 could selectively regulate TGF-β/Smad pathway-associated factors to inhibit activation, migration, and proliferation of HSCs and exert anti-liver fibrosis functions. Exogenous BMP7 has the potential to be used as an anti-liver fibrosis drug.

Mao Mu,1,* Shi Zuo,2,* Rong-Min Wu,3 Kai-Sheng Deng,1 Shuang Lu,1 Juan-Juan Zhu,1 Gao-Liang Zou,1 Jing Yang,1 Ming-Liang Cheng,1 Xue-Ke Zhao1 1Department of Infectious Diseases, The Affiliated Hospital of Guizhou Medical University, Guiyang, Guizhou, China; 2Department of Hepatobiliary Surgery, The Affiliated Hospital of Guizhou Medical University, Guiyang, Guizhou, China; 3Department of Ultrasonography, The Maternity Hospital of Guizhou, Guiyang, Guizhou, China *These authors contributed equally to this work Purpose: Liver fibrosis is a worldwide health issue. Development of effective new drugs for treatment of this disease is of great importance. This study investigated the therapeutic effects of ferulic acid on liver fibrosis invitro and invivo.Materials and methods: Human hepatic stellate cell line (HSC) LX-2 was used for invitro assays. Transforming growth factor β1 (TGF-β1) was used to induce hepatic fibrosis in LX-2 cells. Western blot was used to detect protein levels of collagen I, fibronectin, α-smooth muscle actin (SMA), p-Smad2, p-Smad3, p-p38, and p-JNK. Gene expression was measured by RT-qPCR. Fluorescence staining was used to determine localization of Smad4. CCl4-induced hepatic fibrosis in SD rats was used as an invivo model. Histological features were detected by hematoxylin and eosin staining. Levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), hexadecenoic acid (HA), and hydroxyproline (Hyp) were measured by ELISA.Results: TGF-β1 treatment significantly increased levels of collagen I, fibronectin, α-SMA, p-Smad2, p-Smad3, and Smad4 in LX-2 cells. Ferulic acid improved TGF-β1-induced hepatic fibrosis via regulation of the TGF-β1/Smad pathway. Consistent with invitro data, CCl4 caused severe hepatic fibrosis in SD rats, as determined by ALT, AST, HA, and Hyp upregulation. Protein levels of p-Smad2 and p-Smad3 in liver tissues were significantly increased following treatment with CCl4. All CCL4-induced changes were markedly attenuated by ferulic acid treatment.Conclusion: Ferulic acid potently improved hepatic fibrosis via inhibition of the TGF-β1/Smad pathway invitro and invivo. These findings provided evidence for potential use of ferulic acid to treat or prevent liver fibrosis. Keywords: ferulic acid, TGF-β1, CCl4, hepatic fibrosis, Smad signaling pathway&nbsp

Exergy analysis as well as energy analysis is applied to Chinese comprehensive utilization system, in which oil shale is firstly introduced to fluidized bed (FB) retort for obtaining oil and resulting semicoke is fed to circulating fluidized bed (CFB) reactor for further utilization. During the calculation, linear programming helps optimize the efficiency of FB retort in the system. In the light of the results, the process flow diagram of the whole system is redrawn. Also, this paper discusses how three operating parameters (the retorting temperature, the mass of burned fuel gas and the temperature of circulating ash) influence the system, especially exergy efficiency. With the retorting temperature increasing, the exergy efficiency first increase and then decrease, and the highest exergy efficiency is in the temperature range 460–490 °C. Compared with retorting temperature, the mass of burned fuel gas and circulating ash temperature have less effect on the exergy efficiency of the whole system. But it is a good strategy for stable operation of the system to burn more fuel gas to provide more energy and exergy to FB retorting unit. This work could give further detailed suggestions and more reference data to operate the system stably and efficiently.

Non-alcoholic fatty liver disease (NAFLD) is one of the most prevalent diseases worldwide. Blueberry, combined with probiotics (BP), might be a potential candidate for NAFLD treatment, due to its anti-inflammatory and anti-apoptotic properties. Here, we investigated whether the anti-inflammatory cytokine, IL-22, was involved in the therapeutic process of BP, using cell and rat models of NAFLD. Results indicated that BP significantly reduced lipid droplets and triglyceride (TG) accumulation in NAFLD cells. However, when IL-22 was deficient, the lipid droplets and TG content were significantly increased. In vivo, the serum parameters and pathological degree of NAFLD rats were significantly improved by BP, while IL-22 silencing significantly abolished the BP effect. Immunohistochemistry, immunofluorescence, qRT-PCR, and western blotting showed that the NAFLD group expressed significantly lower levels of IL-22, JAK1, and STAT3, and higher levels of BAX, than the normal group. Furthermore, BP significantly elevated the levels of IL-22, JAK1 and STAT3, and reduced the level of BAX in NAFLD, while IL-22 silencing prevented BP from restoring the expression of JAK1, STAT3, and BAX. We conclude that IL-22 is vital for the therapeutic effect of BP, and acts via activation of JAK1/STAT3 signaling and inhibition of the apoptosis factor BAX, which makes IL-22 a promising target for therapy of NAFLD.

The co-pyrolysis characteristics of oil shale–biomass blends were investigated by open-system pyrolysis with a thermogravimetry–mass spectrometry (TG–MS) analyzer and by closed-system pyrolysis with a fixed-bed reactor. Online Fourier transform infrared (FTIR) spectrometry and gas chromatography–mass spectrometry (GC–MS) were employed to analyze the gas and liquid products that were generated from the closed-system pyrolysis. In addition, the Gaussian program was also employed for quantum chemistry calculations. According to the results, one important reaction mode of oil shale–biomass interactions can be described as the “biogas–kerogen” reaction, which means the combination of kerogen and some free radicals volatilized from the biomass in the temperature range of 180–400 °C. During the co-pyrolysis, the oil shale influences the distribution of the oxygen content in the biomass products, causing an increase of CO2 and CO in the gaseous products and a reduction of oxygen-containing compounds such as alcoh...

Understanding the underlying molecular mechanisms of liver fibrosis is important to develop effective therapy. Herein, we show that focal-adhesion-kinse (FAK) plays a key role in promoting hepatic stellate cells (HSCs) activation in vitro and liver fibrosis progression in vivo. FAK activation is associated with increased expression of α-smooth muscle actin (α-SMA) and collagen in fibrotic live tissues. Transforming growth factor beta-1 (TGF-β1) induces FAK activation in a time and dose dependent manner. FAK activation precedes the α-SMA expression in HSCs. Inhibition of FAK activation blocks the α-SMA and collagen expression, and inhibits the formation of stress fibers in TGF-β1 treated HSCs. Furthermore, inhibition of FAK activation significantly reduces HSC migration and small GTPase activation, and induces apoptotic signaling in TGF-β1 treated HSCs. Importantly, FAK inhibitor attenuates liver fibrosis in vivo and significantly reduces collagen and α-SMA expression in an animal model of liver fibrosis. These data demonstrate that FAK plays an essential role in HSC activation and liver fibrosis progression, and FAK signaling pathway could be a potential target for liver fibrosis.

Purpose Liver fibrosis is a worldwide health issue. Development of effective new drugs for treatment of this disease is of great importance. This study investigated the therapeutic effects of ferulic acid on liver fibrosis in vitro and in vivo. Materials and methods Human hepatic stellate cell line (HSC) LX-2 was used for in vitro assays. Transforming growth factor β1 (TGF-β1) was used to induce hepatic fibrosis in LX-2 cells. Western blot was used to detect protein levels of collagen I, fibronectin, α-smooth muscle actin (SMA), p-Smad2, p-Smad3, p-p38, and p-JNK. Gene expression was measured by RT-qPCR. Fluorescence staining was used to determine localization of Smad4. CCl4-induced hepatic fibrosis in SD rats was used as an in vivo model. Histological features were detected by hematoxylin and eosin staining. Levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), hexadecenoic acid (HA), and hydroxyproline (Hyp) were measured by ELISA. Results TGF-β1 treatment significantly increased levels of collagen I, fibronectin, α-SMA, p-Smad2, p-Smad3, and Smad4 in LX-2 cells. Ferulic acid improved TGF-β1-induced hepatic fibrosis via regulation of the TGF-β1/Smad pathway. Consistent with in vitro data, CCl4 caused severe hepatic fibrosis in SD rats, as determined by ALT, AST, HA, and Hyp upregulation. Protein levels of p-Smad2 and p-Smad3 in liver tissues were significantly increased following treatment with CCl4. All CCL4-induced changes were markedly attenuated by ferulic acid treatment. Conclusion Ferulic acid potently improved hepatic fibrosis via inhibition of the TGF-β1/Smad pathway in vitro and in vivo. These findings provided evidence for potential use of ferulic acid to treat or prevent liver fibrosis.

Liver fibrosis is involved in the progression of most chronic liver diseases. Even though we have made a huge progress in order to understand the pathogenesis of liver fibrosis, however, there is still a lack of productive treatments. Being a traditional Chinese medicine, Platycodin D (PD), an oleanane kind of triterpenoid saponin has been put to extensive use for treating different kinds of illnesses that include not just anti-nociceptive, but also antiviral, anti-inflammatory, and anti-cancer for thousands of years. Nonetheless, there has been no clarification made for its effects on the progression of liver fibrosis. In this manner, we carried out in vitro studies for the purpose of investigating the anti-fibrosis impact of PD. Activation of hepatic stellate cells was evaluated by means of the detection of the proliferation of HSCs and the expression of specific proteins. We discovered the fact that PD had the potential of activating HSCs. Thereafter, we detected the apoptosis and autophagy of the HSCs; as the results suggested, PD induced apoptosis and autophagy of the HSCs. It augmented the expression level of apoptotic proteins that included Bax, Cytochrome C (cyto-c), cleaved caspase3 and cleaved caspase9, in addition to the autophagy relevant proteins, for instance, LC3II, beclin1, Atg5 and Atg9. Further research was carried out for the investigation of the underlying molecular mechanism, and discovered that PD promoted the phosphorylation of JNK and c-Jun. Treating the JNK inhibitor P600125 inhibited the effect of PD, confirming the impact of PD on the regulation of JNK/c-Jun pathway. Thus, we speculated that PD alleviates liver fibrosis and activation of hepatic stellate via promoting phosphorylation of JNK and c-Jun and further altering the autophagy along with apoptosis of HSCs.

Co-pyrolysis of oil shale and organic solid wastes rich in hydrogen is significant for improving shale oil quality and recycling wastes effectively and cleanly. This paper applied thermogravimetric system coupled with mass spectrometry system (TG-MS) to investigate pyrolysis characteristics of low density polyethene (LDPE) and polypropylene (PP), and explore their effects on the pyrolysis behaviors of oil shale during co-pyrolysis. These experiments proved that H radicals and H2 favored the interactions between plastics and oil shale. More specifically, the polymer chain of PP had more side groups than LDPE, and thus PP decomposed more easily and released more H radicals and H2 than LDPE. Accordingly, during the kerogen decomposition, thermogravimetric data indicated that interactions between oil shale and liquid PP were significant, while the interactions between oil shale and liquid LDPE were inconspicuous. However, the data from MS indicated the addition of plastics, especially minor addition of plastic, increased the volatiles of medium molecular weight, which implied that gaseous reactions between volatiles from LDPE and volatiles from oil shale were critical for final product distribution.

Fast co-pyrolysis characteristics of oil shale-wood blends were researched by a bubbling fluidized bed reactor in this paper. An on-line GASMET Fourier transform infrared (FTIR) spectrometer and a gas chromatography-mass spectrometer (GC–MS) were employed for analyzing gas and liquid products. The effect of different blending ratios of oil shale(S)/wood(W) (S:W = 1:0,3:1,1:1,1:3,0:1 in this paper) on the co-pyrolysis products was discussed. The effect of temperature on the characteristics of the co-pyrolysis of S:W = 3:1 was also investigated in this paper. According to the results, the interaction of oil shale and biomass influenced oxygen distribution in volatiles, promoting the generation of CO2 generation and inhibiting the conversion of oxygen-containing compounds like alcohols and acids in pyrolytic oil. The effects of minerals in oil shale and the free radicals generated from wood were concluded according to the experimental results. In addition, as the temperature increased from 430 °C–600 °C, the yield of oil reached maximum at 520 °C with stronger secondary cracking of kerogen and biomass macromolecules.

Background/Aims: Hepatic stellate cells (HSCs) are the primary cell type responsible for liver fibrosis. Our study proved that thymosin beta 4 (Tβ4) has anti-fibrogenic effects in HSCs through PI3K/AKT pathway. However, the underlying mechanisms are not fully elucidated. Circular RNAs (circRNAs) play important roles in fine-tuning gene expression and are often deregulated in cancers. However, the expression profile and clinical significance of in liver fibrosis is still unknown. Therefore, we hypothesize that Tβ4 influences circRNAs in liver fibrosis. Methods: Circular RNA microarray was conducted to identify Tβ4-related circRNAs. Pathway analysis and miRNA response elements analysis was conducted to predict the potential roles of differentially expressed circRNAs in liver fibrosis. CCK8 assays and flow cytometric assays were conducted to clarify the role of circRNA in liver fibrosis. Bioinformatics analysis and in vitro experiments were conducted to clarify the mechanism of circRNA-mediated gene regulation in liver fibrosis. Results: A total of 644 differentially expressed circRNAs were identified between the Tβ4-depleted LX-2 cells and the control LX2 cells. The expression of circRNA-0067835 was significantly increased in the Tβ4-depleted LX-2 cells compared with control. Knockdown of circRNA-0067835 observably decreased LX-2 cell proliferation by causing G1 arrest and promoting apoptosis. Bioinformatics online programs predicted that circRNA-0067835 acted as miR-155 sponge to regulate FOXO3a expression, which was validated using luciferase reporter assay. Conclusion: Our experiments showed that circRNA-0067835 regulated liver fibrosis progression by acting as a sponge of miR-155 to promote FOXO3a expression, indicating that circRNA-0067835 may serve as a potential therapeutic target for patients with liver fibrosis.

Growing evidence has shown that gut microbiome is a key factor involved in liver health. Therefore, gut microbiota modulation with probiotic bacteria, such as Saccharomyces boulardii, constitutes a promising therapy for hepatosis. In this study, we aimed to investigate the protective effects of S. boulardii on D-Galactosamine-induced liver injury in mice. Liver function test and histopathological analysis both suggested that the liver injury can be effectively attenuated by S. boulardii administration. In the meantime, S. boulardii induced dramatic changes in the gut microbial composition. At the phylum level, we found that S. boulardii significantly increased in the relative abundance of Bacteroidetes, and decreased the relative abundance of Firmicutes and Proteobacteria, which may explain the hepatic protective effects of S. boulardii. Taken together, our results demonstrated that S. boulardii administration could change the gut microbiota in mice and alleviate acute liver failure, indicating a potential protective and therapeutic role of S. boulardii.

HBV transgenic mice are used as chronic hepatitis B models for research, but they cannot be infected by HBV to have the hepatitis B as the same as human. In the research, we explored the differences between HBV transgenic mice and normal mice who are injected the concanavalin, A which can injure the livers of the mice. We found some data to show the differences of them after the injection, such as weight, pictures of the liver tissue, alanine transaminase (ALT) and aspartate transaminase (AST) in the blood. The pro-inflammatory cytokines (IL-6, IFN-γ and TNF-α) and T cells (CD4+ cells and CD8+ cells) also showed the livers of HBV transgenic mice were more vulnerable to be hurt than normal mice.

The aim of this study was to investigate the mechanisms of different skin permeability of ibuprofen racemate and enantiomers.The percutaneous permeation of ibuprofen racemate and enantiomers through rabbit normal skin and damaged skin (without stratum corneum [SC]) was investigated in vitro using side-by-side diffusion cells. With the melting temperature-membrane transport model, the flux ratio of enantiomer/racemate was calculated from their thermodynamic properties obtained by differential scanning calorimetry. Attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) study was performed to evaluate the interaction between the enantiomers and the SC. New fluorescent probes were designed and utilized in confocal laser scanning microscopy (CLSM) study for visualization of the enantioselective permeation of the enantiomers through the intact rabbit skin.The flux of (S)-ibuprofen through normal skin was significantly higher than that of (RS)-ibuprofen and (R)-ibuprofen (p0.05), whereas in damaged skin, there was no significant difference (p0.05). The predicted flux ratio of (S)-ibuprofen/(RS)-ibuprofen (2.50) was in close agreement with the experimentally determined ratio (2.48). These results were supported by ATR-FTIR and CLSM studies that indicated that a chiral environment of the skin led to the enantioselective permeation of enantiomers.The chiral nature of the SC and the different physicochemical properties of the enantiomers should be taken into account in the assessment of different skin permeability of the racemate and enantiomers. The synthetic fluorescent probes used in this study could visualize the enantioselective permeation of the chiral compounds across the skin.

A fast half-pixel motion estimation algorithm and its corresponding hardware architecture are presented. Unlike three steps are needed in typical half-pixel motion estimation algorithm, the presented algorithm needs only two steps to obtain all the interpolated pixels of an entire 8 × 8 block. The proposed architecture works in a parallel way and is simulated by Modelsim 6.5 SE, synthesized to the Xilinx Virtex4 XC4VLX15 Field Programmable Gate Array (FPGA) device, and verified by hardware platform. The implementation results show that this architecture can achieve 190 MHz and 11 clock cycles are reduced to complete the entire interpolation process in comparison with typical half-pixel interpolation, which meets the requirements of real-time application for very high defination videos.

[This corrects the article DOI: 10.1371/journal.pone.0201136.].

Objective: To investigate the role of miR-26a-5p in cell proliferation and doxorubicin sensitivity in hepatocellular carcinoma. Methods: We evaluated miR-26a-5p expression in hepatocellular carcinoma tissues and cell lines by reverse transcription polymerase chain reaction. Cell Counting Kit-8 was used to examine cell proliferation. Relationship between miR-26a-5p and aurora kinase A was evaluated by luciferase report system. Western blot was used to detect expression of aurora kinase A. Results: In this study, we observed miR-26a-5p was downregulated in hepatocellular carcinoma tissues and cell lines. Gain-of-function experiments showed that proliferation rate of hepatocellular carcinoma cells decreased under condition of miR-26a-5p mimics. We found miR-26a-5p mimics could enhance doxorubicin sensitivity of hepatocellular carcinoma cells. Further study showed that aurora kinase A was target gene of miR-26a-5p. Suppression of aurora kinase A could lead to lower cell proliferation and higher doxorubicin sensitivity of hepatocellular carcinoma cells. Conclusion: Our study found that miR-26a-5p could inhibit cell proliferation and enhance doxorubicin sensitivity in hepatocellular carcinoma cells by targeting aurora kinase A.

The lipase B from Candida antarctica was subjected to directed evolution suggested by its structure. Mutants of the lipase show significantly increased activity and enantioselectivity toward profen esters compared to the wild type, especially for flurbiprofen ester (ee = 93%, E = 37) and ketoprofen ester (ee = 99%, E > 200).

// Lei Yu 1, 2, * , Bao-fang Zhang 3, * , Ming-liang Cheng 2 , Xue-ke Zhao 2 , Quan Zhang 2 , Ya-xin Hu 2 , Hua-juan Liu 2 , Mao Mu 2 , Bi Wang 2, 4 , Guo-zhen Yang 2 , Li-li Zhu 2 , Shuai Zhang 5 , Yu-mei Yao 2 , Yi-ju Cheng 2 , Wang-sheng Li 6 1 The First Affiliated Hospital of Jinan University, Guangdong 510632, Guangzhou, China 2 The Affiliated Hospital, Guizhou Medical University, Guiyang 550004, Guizhou, China 3 The First Affiliated Hospital, Soochow University, Suzhou 215006, Jiangsu, China 4 Department of Eugenics and Genetics, Guiyang Maternal and Child Health-Care Hospital, Guiyang 550003, Guizhou, China 5 Department of Interventional Radiology, Cancer Hospital of Guizhou Medical University, Guiyang 550004, Guizhou, China 6 Department of Clinical Laboratory, Liupanshui People’s Hospital, Liupanshui 553401, Guizhou, China * These authors contributed equally to this work and should be considered as co-first authors Correspondence to: Ming-liang Cheng, e-mail: gmcmingliang_cheng@163.com Guo-zhen Yang, e-mail: gmcguozhen_yang@163.com Keywords: hepatitis B virus, hepatocellular carcinoma, liver cirrhosis, mutation, risk Received: February 29, 2016 Accepted: April 28, 2016 Published: May 17, 2016 ABSTRACT The long-term outcomes of liver cirrhosis (LC) and hepatocellular carcinoma (HCC) in patients with chronic hepatitis B virus (HBV) infection are associated with specific HBV genotypes and mutations in the virus genome. However, a number of gene-disease association studies have yielded inconsistent results in the field. To investigate this inconsistency, we conducted a meta-analysis from 118 studies involving a total of 9,418 HCC cases, 2,697 LC cases, and 18,785 HBV-infected participants for 11 mutations of HBV to evaluate the epidemiological evidence of the relationship. Overall, 10 mutants (Pre-S mutation, A1762T/G1764A double mutations, G1896A, G1899A, T1753V, C1653T, G1766A, A1762T, G1764A, T1768A) were significantly associated with increased HCC risk with odds ratio (OR) range from 1.80 to 4.27, while no associations were found for C1858T. We found a significant dose–risk relationship between the number of mutations in HBV genome and HCC, in which high risks for HCC were associated with mutation numbers more than 5 (OR = 18.45, 95% CI: 7.86–43.29). By pooling 15 prospective studies, A1762T/G1764A, Pre-S, T1753V, and C1653T mutation was identified as good predictor of HCC risk, showing ORs from 1.73 to 4.54. In addition, significantly elevated LC risks were associated with 6 mutants (A1762T/G1764A double mutations, G1896A, G1899A, T1753V, C1653T, Pre-S mutation), with OR range from 1.76 to 4.10. Our results suggested that HBV mutations alone or in combination may be of clinical significance for predicting hepatocarcinogenesis.

Liver fibrosis results in cirrhosis, liver cancer, and liver failure, which is a major cause of mortality worldwide. Gene therapy is a relatively new paradigm in medicine, with enormous therapeutic potential. The development of an efficient and safe delivery system is essential for clinical gene therapy. In the present study, we evaluated augmenter of liver regeneration/growth factor ERV1-like (ALR/GFER) gene therapeutic effect mediated by a novel minicircle vector (MC-hALR). The results in liver fibrotic rats that received MC-hALR through hydrodynamics-based transfection (HBT) for 8 weeks indicated that the minicircle DNA vector produced a more effective gene therapy effect than traditional plasmids (pcDNA3.1-hALR). Even when we reduced the treatment dose of MC-hALR to 30% (w/w) and the treatment frequency from weekly to biweekly, the in vitro and in vivo results still demonstrated that higher ALR gene expression significantly blocked increases in transforming growth factor-β1 (TGF-β1), platelet derived growth factor-BB (PDGF-BB), and α-smooth muscle aorta (α-SMA) levels; effectively suppressed the production of collagens, especially collagen I; and effectively alleviated liver injury and fibrosis in rats, thereby improving the survival rate of liver fibrotic rats. It is preliminarily concluded that the relative overexpression of MC-hALR inhibits the activation of hepatic stellate cells (HSCs), thereby alleviating liver fibrosis in rats.

Rationale Tumor necrosis factor-alpha (TNF-α) is a potent pro-inflammatory mediator and its expression is up-regulated in chronic obstructive pulmonary disease (COPD). Tristetraprolin (TTP) is implicated in regulation of TNF-α expression; however, whether TTP is involved in cigarette smoke-induced TNF-α expression has not been determined. Methods TTP expression was examined by western blot analysis in murine alveolar macrophages and alveolar epithelial cells challenged without or with cigarette smoke extract (CSE). TNF-α mRNA stability, and the decay of TNF-α mRNA, were determined by real-time quantitative RT-PCR. TNF-α protein levels were examined at the same time in these cells. To identify the molecular mechanism involved, a construct expressing the human beta-globin reporter mRNA containing the TNF-α 3’-untranslated region was generated to characterize the TTP targeted site within TNF-α mRNA. Results CSE induced TTP down-regulation in alveolar macrophages and alveolar epithelial cells. Reduced TTP expression resulted in significantly increased TNF-α mRNA stability. Importantly, increased TNF-α mRNA stability due to impaired TTP function resulted in significantly increased TNF-α levels in these cells. Forced TTP expression abrogated the increased TNF-α mRNA stability and expression induced by CSE. By using the globin reporter construct containing TNF-α mRNA 3’-untranslated region, the data indicate that TTP directly targets the adenine- and uridine-rich region (ARE) of TNF-α mRNA and negatively regulates TNF-α expression at the post-transcriptional level. Conclusion The data demonstrate that cigarette smoke exposure reduces TTP expression and impairs TTP function, resulting in significantly increased TNF-α mRNA stability and excessive TNF-α expression in alveolar macrophages and epithelial cells. The data suggest that TTP is a novel post-transcriptional regulator and limits excessive TNF-α expression and inflammatory response induced by cigarette smoke.

Lung fibrosis is a major medical problem for the aging population worldwide. Fibroblast migration plays an important role in fibrosis. Focal Adhesion Kinase (FAK) senses the extracellular stimuli and initiates signaling cascades that promote cell migration. This study first examined the dose and time responses of FAK activation in human lung fibroblasts treated with platelet derived growth factor BB (PDGF-BB). The data indicate that FAK is directly recruited by integrin β1 and the subsequent FAK activation is required for fibroblast migration on fibronectin. In addition, the study has identified that α5β1 and α4β1 are the major integrins for FAK-mediated fibroblast migration on fibronect. In contrast, integrins αvβ3, αvβ6 and αvβ8 play a minor but distinct role in fibroblast migration on fibronectin. FAK inhibitor significantly reduces PDGF-BB stimulated fibroblast migration. Importantly, FAK inhibitor protects bleomycin-induced lung fibrosis in mice. FAK inhibitor blocks FAK activation and significantly reduces signaling cascade of fibroblast migration in bleomycin-challenged mice. Furthermore, FAK inhibitor decreases lung fibrotic score, collagen accumulation, fibronectin production and myofibroblast differentiation in in bleomycin-challenged mice. These data demonstrate that FAK mediates fibroblast migration mainly via integrin β1. Furthermore, the findings suggest that targeting FAK signaling is an effective therapeutic strategy against fibrosis.

Augmenter of liver regeneration (ALR), produced and released by hepatocytes, has cytoprotective and immunoregulatory effects on liver injury, and has been used in many experimental applications. However, little attention has been paid to the effects of ALR on concanavalin A (Con A)-induced hepatitis. The purpose of this paper is to explore the protective effect of ALR on Con A-induced hepatitis and elucidate potential mechanisms. We found that the ALR pretreatment evidently reduced the amount of ALT and AST in serum. In addition, pro-inflammatory cytokines, chemokines and iNOS were suppressed. ALR pretreatment also decreased CD4(+), CD8(+) T cell infiltration in liver. Besides, we observed that ALR pretreatment was capable of suppressing the activation of several signaling pathways in Con A-induced hepatitis. These findings suggest that ALR can obviously weaken Con A-induced hepatitis and ALR has some certain immune regulation function.

Cyclophosphamide (CTX) has immunosuppressive effects and has been wildly used as one anti-cancer drug in clinical. Significant toxicity has been noticed particularly in the reproductive system. CTX promotes the maturation of ovarian follicles, decreases follicular reserve, and ultimately lead to ovarian failure or even premature ovarian failure (POF). The placental extract (HPE) has been shown to have some beneficial impact on reproductive system; however, little is known regarding to the effect of HPE on protecting CTX-induced ovarian injury and the mechanism involved. Whether human placental extracts (HPE) has a protective effect on CTX-induced toxicity on ovarian was studied by using a CTX-induced ovarian injury animal model. The effects of HEP on histopathology, the number of atretic follicles, the weight of the ovary, serum hormone levels, and apoptosis in granulosa cells were studied in mice with CTX or control vehicle. Our results have demonstrated that HPE inhibited p-Rictor, reduced the expression of Bad, Bax and PPAR, and activated Akt and Foxo3a (increased their phosphorylation). Mice treated with HPE showed higher ovarian weight, lower number of atretic follicles, higher serum levels of the hormones E2 and progesterone, and lower apoptosis and serum levels of LH and FSH in granulosa cells, than that in the control animal group. Our data show that ovarian injury can be attenuated by HPE. HPE likely protects follicular granulosa cells from undergoing significant apoptosis and reduce atresia follicle formation, therefore, alleviates CTX-induced ovarian injury.

Polypropylene (PP)/ethylene-octene copolymer (POE) blends with 10–50wt% POE composition were prepared using a twin-screw extruder in the melt state. Mechanical properties of PP and PP/POE blends were tested and the effect of POE content on the crystalline morphology and structure, melting and crystallization behavior, compatiblilty, phase morphology, and the interface cohesiveness of the blends were investigated by polarizing optical microscope (POM), wide-angle X-ray diffraction (WAXD), differential scanning calorimetry (DSC), dynamic mechanical thermal analysis (DMTA), and scanning electron microscopy (SEM). The relationship between mechanical properties and microstructure of the PP/POE blends is discussed. The results showed that POE had a dual function of both reinforcing and toughening PP in the range from 10–40wt%, which was attributed to the integrated functions of the degree of crystallinity of the PP phase, phase morphology, and interface cohesiveness of the blend.

Hepatitis B virus (HBV) infection is the predominant risk factor for chronic hepatitis B (CHB), liver cirrhosis (LC) and hepatocellular carcinoma (HCC). Recently, genome-wide association studies have identified human leukocyte antigen (HLA)-DP polymorphisms (rs3077 and rs9277535) as a new chronic HBV infection susceptibility locus. Since then, the relationship between HLA-DP polymorphisms and various outcomes of HBV infection has been reported. However, the results have been inconclusive. To derive a more precise estimation of the relationship between HLA-DP polymorphisms and various outcomes of HBV infection, a meta-analysis of 62,050 subjects from 29 case-control studies was performed. We found that rs3077 and rs9277535 in HLA-DP significantly decreased HBV infection risks and increased HBV clearance possibility in a dose-dependent manner. In the subgroup analysis by ethnicity, study design and sample size, significant associations were found for these polymorphisms in almost all comparisons. Meanwhile, haplotype analyses of the two polymorphisms revealed a significant association between the combination of these alleles and HBV infection outcomes. However, no significant results were observed in HCC development. Our results further confirm that genetic variants in the HLA-DP locus are strongly associated with reduced HBV infection and increased the likelihood of spontaneous viral clearance.

To investigate the clinical features and rate of natural viral clearance in patients with hepatitis C virus (HCV) infection acquired by blood transfusion from a single donor.Ninety-six patients who acquired HCV infection between January 1998 and December 2002, upon receipt of donated blood from a single infected individual in Guizhou,were included in this retrospective cross-sectional study. Patients were clinically assessed to determine levels of anti-HCV antibodies, HCV RNA and biochemical indicators of liver function,as well as features of liver structure (by abdominal B ultrasonography and elastography). HCV genetic testing was used to determine the virus genotype. Measurement data were expressed as mean ± standard deviation. Count data were analyzed by the x² test,with P less than 0.05 indicating statistical significance.All 96 patients tested positive for antiHCV antibodies. The majority of patients (70%; 34:33 male:female) had HCV RNA more than or equal to 1.0 * 103 copies/ml. All patients carried the same HCV genotype as the single blood donor:genotype lb. The overall rate of natural HCV clearance was 30.2%. but males had a significantly lower rate (19.0% (8/42) vs. females:38.9% (21/54);x²=4.41,P=0.023) as did older patients (more than 40 years-old:16.1% (5/31) vs .less than or equal to 40 years-old:36.9% (24/65);x²=4.30,P=0.028). The overall rate of chronic HCV infection (CHC) was 69.8%,but the rate was significantly lower in younger patients (less than or equal to 40 years-old:63.1% (41/65) vs. more than 40 years-old:83.9% (26/31);x²=6.67,P=0.028). Among the 67 patients with CHC,12 had symptoms of mild weakness,anorexia and abdominal distention,11 had elevated serum alanine aminotransferase (116.25 +/- 24.65 U/L) and stage 3 or 4 fibrosis (liver elasticity values more than or equal to 5.1 kPa),and three had mildly abnormal serum bilirubin (32.56 ± 5.28 mumol/L). Fifteen patients showed signs of chronic hepatitis and one patient showed signs of cirrhosis by abdominal B ultrasonography. None of the patients showed signs of hepatocellular carcinoma.The course of blood transfusion acquired HCV infection is largely unknown and natural viral clearance rate may be associated with sex-and age-related factors.

To investigate the regulative mechanism of the diterpene phenol extract of Rosmarinus Officinalis (DERO) on the imbalance of collagen metabolism of the lung tissue in pulmonary fibrosis rats.Fifty healthy Sprague-Dawley rats were randomly divided into the normal saline group (NS), the bleomycin-induced lung injury group (BLM), the low dose DERO group (at the daily dose of 50 mg/kg), the moderate dose DERO group (at the daily dose of 100 mg/kg), and the high dose DERO group (at the daily dose of 200 mg/kg), 10 in each group (abbreviated as DERO 1, 2, 3, respectively). The pulmonary fibrosis rat model was prepared by disposable intratracheal instillation of bleomycin. DERO was administered by gastrogavage as intervention during the repairing process of lung injury. On the morning of the 29th day, the rats' lung tissue was extracted. The karyocyte number, collagen protein, type I collagen (collagen I) and transforming growth factor-beta type II receptor (TGFbetaR II), Smad4 mRNA expressions were semi-quantitatively determined using tissue microarray, HE staining, collagen fiber dyeing, immunohistochemical assay, and in situ hybridization. Using real-time fluorescent quantification RT-PCR, the mRNA expression of transforming growth factor-beta1 (TGF-beta1) were detected.Compared with the NS group, the collagen deposition of the lung tissue was obvious and the inflammatory infiltration was more severe in the BLM group (P0.05, P0.01). There was no statistical difference in the aforesaid 4 indices between the DERO1 group and the BLM group (P0.05). The collagen deposition and the inflammatory infiltration were obviously alleviated in the DERO2 and DERO3 groups (P0.05, P0.01). Compared with the NS group, the mRNA expressions of collagen-I, TGF-beta1 R II, Smad4, and TGF-beta1 were obviously up-regulated in the BLM group (P0.05, P0.01). Compared with the BLM group, the aforesaid four indices were not statistically changed in the DERO1 group (P0.05). But the mRNA expressions of collagen-I, TGF-beta1 R II, Smad4, and TGF-beta1 were obviously downregulated in the DERO2 and DERO3 groups (P0.05, P0.01). But the down-regulation of Smad4 expression was not obvious in the DERO2 and the DERO3 groups (P0.05). Compared with the DERO1 group, the mRNA expressions of collagen-I, TGF-beta1, R II, TGFbeta1 were all obviously lower in the DERO2 and the DERO3 groups (P0.05). But there was no statistical difference in the aforesaid 4 indices between the DERO2 group and the DERO3 group (P0.05).DERO could regulate imbalanced collagen metabolism of pulmonary fibrosis. It could inhibit excessive deposition of collagen fibers, especially excessive deposition of collagen- I. Its mechanisms might be realized by inhibiting up-regulation of TGF-beta1 and TGFbetaR II mRNA expressions, thus interfering the activation of TGF-beta-Smad signaling pathway on target genes, especially on type I procollagen target gene.

Two ketoreductases from Candida glabrata were used for the asymmetric reduction of prochiral substituted acetophenones displayed different enantiopreference toward para-, meta-substituted and ortho-halogen substituted acetophenones with excellent enantioselectivity. Homology modeling and docking analysis were in conformity with this interested enantiopreference obtained from experimental tests. The reduction of a series of other substituted aryl ketones was also investigated using the two ketoreductases.