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Artificial multienzyme biocatalysts have played a crucial role in biosynthesis because they allow for conducting complex reactions. Here, we reorted a facile approach to fabricate multienzyme nanodevices with high catalytic activity and stability based on protein assembly and chemical crosslinking. The self-assembled partner SpyCatcher and SpyTag were genetically fused with 2,3-butanediol hydrogenase and formate hydrogenase to produce KgBDH-SC (SpyCatcher-fused 2,3-butanediol hydrogenase) and FDH-ST (SpyTag-fused formate hydrogenase), respectively. After assembling the two fusion proteins, the complexes were then immobilized on the functionalized silicon dioxide nanoparticles by glutaraldehyde, forming KgBDH-SC-ST-FDH-SiO 2 with the capability of reducing 2-hydroxyacetophenone to (R)-1-phenyl-1,2-ethanediol with cofactor regeneration. Under the optimal conditions, the created co-immobilized enzymes performed 49% activity recovery compared with the mixture of free enzymes as well as showed 2.9-fold higher catalytic activity than the traditional random co-immobilized enzymes. Moreover, KgBDH-SC-ST-FDH-SiO 2 showed better pH stability and organic solvents stability than the free enzymes, and remained 52.5% overall catalytic activity after 8 cycles. Finally, the co-immobilized enzymes can reduce 60 mM HAP for fabrication of (R)-PED with cofactor regeneration in the phosphate buffer reaction system, affording 83.9% yield and above 99% optical purity., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Objective: Despite the remarkable progress in efforts to control disease spread, the nationwide elimination of hepatitis B in China is still hindered by the persistently high rate of hepatitis B virus (HBV) infection in Western China. This study aimed to evaluate the strategy of hepatitis B prevention and control in Western China and identify potential areas and strategies for improvement., Methods: Susceptible population vaccination, health education, professional training of doctors, and other prevention and control measures have been implemented in Wuwei city since 2010. Data were obtained from three representative cross-sectional serosurveys conducted in 2010, 2013, and 2015. The serum samples were subjected to enzyme-linked immunosorbent assays to detect the following seromarkers: HBV surface antigen (HBsAg), antibody against hepatitis B surface antigen (anti-HBs), and antibody against hepatitis B core antigen (anti-HBc). Estimates of variance were determined using Taylor series linearization methods., Results: The three serosurveys revealed decreases in the prevalence of HBsAg (7.19% in 2010 vs. 6.51% in 2013 vs. 5.87% in 2015) and anti-HBc positivity (43.89% vs. 32.87% vs. 28.46%) and an increase in the prevalence of anti-HBs positivity (49.07% vs. 53.66% vs. 53.72%) over time. From 2010 to 2015, the legally reported incidence of hepatitis B in Wuwei city decreased from 686.53/100,000 to 53.72/100,000. Notably, persistently high HBsAg-positive rates (above 5.40%) were observed among subjects aged 20-69 years old in the three serosurveys; the prevalence of HBsAg was above 1% among children younger than 10 years old. Furthermore, rural subjects had higher rates of HBsAg and anti-HBc positivity than their urban counterparts (6.04% vs. 4.83% and 30.26% vs. 20.35%, respectively) in 2015 but had a lower rate of anti-HBs positivity (49.68 vs. 55.18%). Multivariate regression analysis showed that age, urban and rural areas, and education level were the main factors affecting HBV infection., Conclusion: Although vaccine-based prevention and control measures reduced the rate of HBV infection in Wuwei City over time, the hepatitis B infection rate in children younger than 10 years was still higher than the national average level. Therefore, the prevention and control of mother-to-child transmission and the management of the infected should be the focus of future prevention and control work., (Copyright © 2020 The Editorial Board of Biomedical and Environmental Sciences. Published by China CDC. All rights reserved.)

Objectives: To explore changes in the quality of life(QoL)in patients with non-severe aplastic anemia(NSAA)after 2 years of cyclosporine A(CsA)therapy, and possible factors may affect the QoL. Methods: Patients with de novo NSAA from January 2014 to 2016 who had been treated with only CsA for at least 2 years in the outpatient department of Peking Union Medical College Hospital were instructed to fill-in the SF-36 form before and after 2 years of CsA treatment. Data from NSAA were compared with those of normal controls; patients' information such as age, sex, education, annual income, type of payment, and compliance were collected, disease severity and response to treatment were also evaluated. Results: A total of 52 patients were included in our study with 27(51.9%)men and 25(48.1%)women, with the medium age of 48(21-85)years. After 2 years of treatment, 15(28.8%)patients achieved complete response(CR), 25(48.1%)achieved partial response(PR), and 12(23.1%)patients had no response(NR). The overall response rate(ORR)was 76.9%. Before the therapy, SF-36 scores in patients with NSAA were significantly lower than that of normal controls either in physical or mental component summaries( P <0.05). However, after 2 years of therapy, patients with NSAA had significant improvement of mental component summaries and recovered to normal with even higher scores in mental health(MH)(65.9±17.6 vs 59.7±22.9, P =0.014)and energy/vitality(VT)(58.8±20.1 vs 52.3±20.9, P =0.023)compared with normal controls, although they still had comparatively lower scores in physical component summaries. No associations were found between QoL and age, sex, educational level, family income, type of payment, patient adherence, or transfusion dependency. Patients with higher ECOG (the Eastern Cooperative Oncology Group score)at the beginning experienced greater progress in QoL compared to those with lower ECOG. Both patients with CR and PR had shown significant improvement in QoL. Conclusion: Patients with NSAA had impaired QoL compared with normal patients. CsA treatment can improve the QoL, especially in mental component summaries. Patients can benefit from the treatment regardless of their social status, and patients with lower ECOG at the beginning seem to benefit more from the therapy.

Although the prevalence of brucellosis in Inner Mongolia Autonomous Region currently remains high, data available on the epidemiological of circulating Brucella abortus strains were limited. A total of 75 isolates obtained from cattle, sheep, and humans were analysed using both the classical method and multiple locus variable-number tandem repeat analysis (MLVA). There are at least three B. abortus biovars (1, 3 and 6) in this region, and B. abortus biovar 3 is the predominant one. Ten known MLVA-11 genotypes were identified, of which five genotypes (72, 75, 78, 82 and 210) were shared among strains from this study and others previously collected in two to seven different nations, suggesting that this population has multiple geographic origins. An MLVA-16 assay sorted the 75 B. abortus strains into two groups (I and II), 5 clusters (A-E) and 44 genotypes (GT1-44), with 26 unique genotypes represented by single isolates, indicating that these B. abortus brucellosis cases were not directly epidemiologically related. The remaining 18 shared genotypes (among a total of 47 isolates) were represented by two to eight isolates, suggesting that there were epidemiologically related pathogens from each shared genotype among the cases. Importantly, the cluster B1 branch including 22 cluster isolates with identical or similar genotypes confirmed the occurrence of a concentrated outbreak epidemic in the eastern region during 1988-1995. This work will contribute to better understanding of B. abortus brucellosis epidemiology in Inner Mongolia., (Copyright © 2020. Published by Elsevier B.V.)

Microbial contamination, especially in large-scale processes, is partly a life-or-death issue for industrial fermentation. Therefore, the aim of this research was to create an antimicrobial contamination system in Bacillus subtilis 168 (an ideal acetoin producer for its safety and acetoin synthesis potential). First, introduction of the formamidase (FmdA) from Helicobacter pylori and the phosphite dehydrogenase (PtxD) from Pseudomonas stutzeri enabled the engineered Bacillus subtilis to simultaneously assimilate formamide and phosphite as nitrogen (N) and phosphorus (P) sources. Thus, the engineered B. subtilis became the dominant population in a potentially contaminated system, while contaminated microbes were starved of key nutrients. Second, stepwise metabolic engineering via chromosome-based overexpression of the relevant glycolysis and acetoin biosynthesis genes led to a 1.12-fold increment in acetoin titer compared with the starting host. Finally, with our best acetoin producer, 25.56 g/L acetoin was synthesized in the fed-batch fermentation, with a productivity of 0.33 g/L/h and a yield of 0.37 g/g under a nonsterilized and antibiotic-free system. More importantly, our work fulfills many key criteria of sustainable chemistry since sterilization is abolished, contributing to the simplified fermentation operation with lower energy consumption and cost.

Central nervous system leukemia (CNSL) relapse is relatively common among Philadelphia chromosome-positive (Ph+) leukemia patients who undergo allogeneic hematopoietic stem cell transplantation (allo-HSCT). The prognosis of patients is dismal for those with a BCR-ABL T315I mutation, which is resistant to TKIs including second-generation drugs. We assessed ponatinib for nine patients with recurrent Ph+ CNSL and a T315I mutation after allo-HSCT, including five patients with Ph+ acute lymphoblastic leukemia and four with chronic myelogenous leukemia. Five patients experienced isolated CNSL relapse, and four experienced CNSL with hematologic relapse. All patients received ponatinib combined with intrathecal chemotherapy, and four patients with hematologic relapse received systemic chemotherapy and/or donor lymphocyte infusion. All patients achieved a deep molecular response and central nervous system remission (CNSR) at a median time of 1.5 (range: 0.7-3) months after ponatinib treatment. Two patients experienced a second CNSL relapse due to ponatinib reduction, but they achieved CNSR again after an increase to the standard dosage. Six patients developed graft versus host disease. By April 1, 2019, eight patients were alive, and one died of pneumonia. The median time of survival after the first CNSL relapse posttransplantation was 18 (range: 11.2-48.5) months. Our data from a small number of samples suggests that ponatinib is effective for recurrent Ph+ CNSL patients with a BCR-ABL T315I mutation after allo-HSCT and warrants broader clinical evaluation., (© 2019 UICC.)

Montmorillonite (MMT) presents nonocclusive lamellar structure which restricts the potential use for sustained drug release. To solve the limitation, the quaternized pectin (QP) was synthesized and firstly introduced to form QP-MMT hybrid film containing 5-FU. The Fourier transform infrared spectroscopy (FT-IR) and X-Ray diffraction (XRD) were employed to determine the variation of the functional group and crystallinity between pectin and QP. The resultant composite film was characterized by FT-IR, XRD and Field Emission Scanning Electron Microscope. The results of the characterization indicated that intercalation reaction happened in the blending process. The optimum film showed high value of drug encapsulation efficiency (36.50%) and loading efficiency (80.30%). The in vitro drug release studies revealed that the MMT significantly improved the sustained-release performance in all simulated mediums. The cumulative release rate of sample QP 10 -MMT 0.1 was all around 20% in the first half-hour in all simulated mediums and sustained increased for more than 8 h. The cytotoxicity assay was performed to prove the great biocompatibility of QP-MMT hybrid film. The present study introduced a facile route to prepare the composite film which presented sustained drug release performance., (Copyright © 2020. Published by Elsevier B.V.)

Background and Aims: DNA damage-induced NF-κB activation is a major obstacle to effective antitumour chemotherapy. Long noncoding RNAs (lncRNAs) that regulate chemoresistance of cancer cells remain largely unknown. This study aimed to characterize the lncRNAs that may affect chemotherapy sensitivity., Approach and Results: We found that lncRNA PDIA3P1 (protein disulfide isomerase family A member 3 pseudogene 1) was up-regulated in multiple cancer types and following treatment with DNA-damaging chemotherapeutic agents, like doxorubicin (Dox). Higher PDIA3P1 level was associated with poorer recurrence-free survival of human hepatocellular carcinoma (HCC). Both gain-of-function and loss-of-function studies revealed that PDIA3P1 protected cancer cells from Dox-induced apoptosis and allowed tumor xenografts to grow faster and to be more resistant to Dox treatment. Mechanistically, miR-125a/b and miR-124 suppressed the expression of tumor necrosis factor receptor-associated factor 6 (TRAF6), but PDIA3P1 bound to miR-125a/b/miR-124 and relieved their repression on TRAF6, leading to activation of the nuclear factor kappa B (NF-κB) pathway. Consistently, the effect of PDIA3P1 inhibition in promoting Dox-triggered apoptosis was antagonized by silencing the inhibitor of κBα (IκBα) or overexpressing TRAF6. Administration of BAY 11-7085, an NF-κB inhibitor attenuated PDIA3P1-induced resistance to Dox treatment in mouse xenografts. Moreover, up-regulation of PDIA3P1 was significantly correlated with elevation of TRAF6, phosphorylated p65, or NF-κB downstream anti-apoptosis genes in human HCC tissues. These data indicate that enhanced PDIA3P1 expression may confer chemoresistance by acting as a microRNA sponge to increase TRAF6 expression and augment NF-κB signaling. Subsequent investigations into the mechanisms of PDIA3P1 up-regulation revealed that human homologue of mRNA transport mutant 4 (hMTR4), which promotes RNA degradation, could bind to PDIA3P1, and this interaction was disrupted by Dox treatment. Overexpression of hMTR4 attenuated Dox-induced elevation of PDIA3P1, whereas silencing hMTR4 increased PDIA3P1 level, suggesting that Dox may up-regulate PDIA3P1 by abrogating the hMTR4-mediated PDIA3P1 degradation., Conclusion: There exists a hMTR4-PDIA3P1-miR-125/124-TRAF6 regulatory axis that regulates NF-κB signaling and chemoresistance, which may be exploited for anticancer therapy., (© 2019 The Authors. Hepatology published by Wiley Periodicals, Inc., on behalf of American Association for the Study of Liver Diseases.)

Cold-adapted pullulanase with high catalytic activity and stability is of special interest for its wide application in cold starch hydrolysis, but few pullulanases displaying excellent characteristics at ambient temperature and acidic pH have hitherto been reported. Here, a novel pullulanase from Bacillus methanolicus PB1 was successfully expressed in Escherichia coli BL21 (DE3) and determined to be a cold-adapted type I pullulanase (PulPB1) with maximum activity at 50 °C and pH 5.5. The recombinant PulPB1 showed great stability, its half-life at 50 °C was 137 h. PulPB1 can efficiently hydrolyze pullulan and amylopectin, with activities of 292 and 184 U/mg at 50 °C and pH 5.5, respectively. Moreover, the N-terminal domain of PulPB1 was found to significantly affect the enzymatic performance. Following truncation of the N-terminal domain, the activity towards pullulan decreased markedly from 292 to 141 U/mg and the half-life at 50 °C decreased from 137 to 10 h. Compared to the hydrolysis system with amyloglucosidase alone, the catalytic efficiency showed a 2.4-fold increase on combining PulPB1 with amyloglucosidase for amylopectin hydrolysis at 40 °C. This demonstrates that PulPB1 is promising for development as a superior candidate for cold amylopectin hydrolysis., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

As a new wastewater biological nitrogen removal process, anammox and partial denitrification coupling not only plays a significant role in the nitrogen cycle, but also holds high engineering application value. Because anammox and some denitrifying bacteria are coupled under harsh living conditions, certain operating conditions and mechanisms of the coupling process are not clear; thus, it is more difficult to control the process, which is why the process has not been widely applied. This paper analyzes the research focusing on the coupling process in recent years, including anammox and partial denitrification coupling process inhibitors such as nitrogen (NH 4 + , NO 2 - ), organics (toxic and non-toxic organics), and salts. The mechanism of substrate removal in anammox and partial denitrification coupling nitrogen removal is described in detail. Due to the differences in process methods, experimental conditions, and sludge choices between the rapid start-up and stable operation stages of the reactor, there are significant differences in substrate inhibition. Multiple process parameters (such as pH, temperature, dissolved oxygen, redox potential, carbon-to-nitrogen ratio, and sludge) can be adjusted to improve the coupling of anammox and partial denitrification to modify nitrogen removal performance., Competing Interests: There are no conflicts of interest to declare., (This journal is © The Royal Society of Chemistry.)

Epigallocatechin gallate (EGCG), a major polyphenol in green tea, exhibits diverse biological activities. Previous studies show that EGCG could effectively suppress HBV gene expression and replication, but the role of EGCG in HBV replication and its underlying mechanisms, especially the signaling pathways involved, remain unclear. In this study we investigated the mechanisms underlying EGCG inhibition on HBV replication with a focus on the signaling pathways. We showed that EGCG (12.5-50 μM) dose-dependently inhibited HBV gene expression and replication in HepG2.2.15 cells. Similar results were observed in HBV mice receiving EGCG (25 mg· kg -1 · d -1 , ip) for 5 days. In HepG2.2.15 cells, we showed that EGCG (12.5-50 μM) significantly activate ERK1/2 MAPK signaling, slightly activate p38 MAPK and JAK2/STAT3 signaling, while had no significant effect on the activation of JNK MAPK, PI3K/AKT/mTOR and NF-κB signaling. By using specific inhibitors of these signaling pathways, we demonstrated that ERK1/2 signaling pathway, but not other signaling pathways, was involved in EGCG-mediated inhibition of HBV transcription and replication. Furthermore, we showed that EGCG treatment dose-dependently decreased the expression of hepatocyte nuclear factor 4α (HNF4α) both at the mRNA and protein levels, which could be reversed by pretreatment with the ERK1/2 inhibitor PD98059 (20 μM). Moreover, we revealed that EGCG treatment dose-dependently inhibited the activity of HBV core promoter and the following HBV replication. In summary, our results demonstrate that EGCG inhibits HBV gene expression and replication, which involves ERK1/2-mediated downregulation of HNF4α.These data reveal a novel mechanism for EGCG to inhibit HBV gene expression and replication.

RNA-binding proteins (RBPs) play important roles in regulating the expression of genes involved in human physiological and pathological processes, especially in cancers. Many RBPs have been found to be dysregulated in cancers; however, there was no tool to incorporate high-throughput data from different dimensions to systematically identify cancer-related RBPs and to explore their causes of abnormality and their potential functions. Therefore, we developed a database named RBPTD to identify cancer-related RBPs in humans and systematically explore their functions and abnormalities by integrating different types of data, including gene expression profiles, prognosis data and DNA copy number variation (CNV), among 28 cancers. We found a total of 454 significantly differentially expressed RBPs, 1970 RBPs with significant prognostic value, and 53 dysregulated RBPs correlated with CNV abnormality. Functions of 26 cancer-related RBPs were explored by analysing high-throughput RNA sequencing data obtained by crosslinking immunoprecipitation, and the remaining RBP functions were predicted by calculating their correlation coefficient with other genes. Finally, we developed the RBPTD for users to explore functions and abnormalities of cancer-related RBPs to improve our understanding of their roles in tumorigenesis. Database URL: http: //www.rbptd.com., (© The Author(s) 2020. Published by Oxford University Press.)

Considerable attention has been given to the development of robust fermentation processes, but microbial contamination and phage infection remain deadly threats that need to be addressed. In this study, a robust Escherichia coli BL21(DE3) strain was successfully constructed by simultaneously introducing a nitrogen and phosphorus (N&P) system in combination with a CRISPR/Cas9 system. The N&P metabolic pathways were able to express formamidase and phosphite dehydrogenase in the host cell, thus enabled cell growth in auxotrophic 3-(N-morpholino)propanesulfonic acid medium with formamide and phosphite as nitrogen and phosphorus sources, respectively. N&P metabolic pathways also allowed efficient expression of heterologous proteins, such as green fluorescent protein (GFP) and chitinase, while contaminating bacteria or yeast species could hardly survive in this medium. The host strain was further engineered by exploiting the CRISPR/Cas9 system to enhance the resistance against phage attack. The resultant strain was able to grow in the presence of T7 phage at a concentration of up to 2 × 10 7 plaque-forming units/ml and produce GFP with a yield of up to 30 μg/10 9 colony-forming units, exhibiting significant advantages over conventional engineered E. coli. This newly engineered, robust E. coli BL21(DE3) strain therefore shows great potential for future applications in industrial fermentation., (© 2019 Wiley Periodicals, Inc.)

Micropeptides (≤100 amino acids) are essential regulators of physiological and pathological processes, which can be encoded by small open reading frames (smORFs) derived from long non-coding RNAs (lncRNAs). Recently, lncRNA-encoded micropeptides have been shown to have essential roles in tumorigenesis. Since translated smORF identification remains technically challenging, little is known of their pathological functions in cancer. Therefore, we created classifiers to identify translated smORFs derived from lncRNAs based on ribosome-protected fragment sequencing and machine learning methods. In total, 537 putative translated smORFs were identified and the coding potential of five smORFs was experimentally validated via green fluorescent protein-tagged protein generation and mass spectrometry. After analyzing 11 lncRNA expression profiles of seven cancer types, we identified one validated translated lncRNA, ZFAS1, which was significantly up-regulated in hepatocellular carcinoma (HCC). Functional studies revealed that ZFAS1 can promote cancer cell migration by elevating intracellular reactive oxygen species production by inhibiting nicotinamide adenine dinucleotide dehydrogenase expression, indicating that translated ZFAS1 may be an essential oncogene in the progression of HCC. In this study, we systematically identified translated smORFs derived from lncRNAs and explored their potential pathological functions in cancer to improve our comprehensive understanding of the building blocks of living systems., (Copyright © 2019 Guo, Meng, Zhai, Xie, Zhao, Li, Zhou, Li, Liu, Yang and Wu.)

Background: The significance of multi-site phosphorylation of BCL-2 protein in the flexible loop domain remains controversial, in part due to the lack of structural biology studies of phosphorylated BCL-2., Objective: The purpose of the study is to explore the phosphorylation induced structural changes of BCL-2 protein., Methods: We constructed a phosphomietic mutant BCL-2(62-206) (t69e, s70e and s87e) (EEEBCL- 2-EK (62-206)), in which the BH4 domain and the part of loop region was truncated (residues 2-61) to enable a backbone resonance assignment. The phosphorylation-induced structural change was visualized by overlapping a well dispersed 15N-1H heteronuclear single quantum coherence (HSQC) NMR spectroscopy between EEE-BCL-2-EK (62-206) and BCL-2., Results: The EEE-BCL-2-EK (62-206) protein reproduced the biochemical and cellular activity of the native phosphorylated BCL-2 (pBCL-2), which was distinct from non-phosphorylated BCL-2 (npBCL-2) protein. Some residues in BH3 binding groove occurred chemical shift in the EEEBCL- 2-EK (62-206) spectrum, indicating that the phosphorylation in the loop region induces a structural change of active site., Conclusion: The phosphorylation of BCL-2 induced structural change in BH3 binding groove., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Glucaric acid (GA) is a major value-added chemicals feedstock and additive, especially in the food, cosmetics, and pharmaceutical industries. The increasing demand for GA is driving the search for a more efficient and less costly production pathway. In this study, a new in vitro multi-enzyme cascade system was developed, which converts sucrose efficiently to GA in a single vessel. The in vitro system, which does not require adenosine triphosphate (ATP) or nicotinamide adenine dinucleotide (NAD + ) supplementation, contains seven enzymes. All enzymes were chosen from the BRENDA and NCBI databases and were expressed efficiently in Escherichia coli BL21(DE3). All seven enzymes were combined in an in vitro cascade system, and the reaction conditions were optimized. Under the optimized conditions, the in vitro seven-enzyme cascade system converted 50 mm sucrose to 34.8 mm GA with high efficiency (75 % of the theoretical yield). This system represents an alternative pathway for more efficient and less costly production of GA, which could be adapted for the synthesis of other value-added chemicals., (© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

A rare and novel Cr-substituted polyoxoniobate (PONb), [Cr 2.5 Nb 27.5 O 66 (OH) 20 (H 2 O) 2 ] 7- , has been synthesized by a combination of hydrothermal and conventional solution methods. The PONb shows an unknown tetrameric structure, which is the largest Cr-containing PONb to date. Interestingly, every two PONb tetramers can be joined together by alkali-metal cations to form a discrete cubelike ionic cluster. The obtained PONb not only enriches the very limited members of the Cr-substituted PONb family but also exhibits high ionic conductivity.

Nonalcoholic fatty liver disease (NAFLD) is a very common chronic hepatic disease, with nonalcoholic steatohepatitis (NASH) as a major and severe subcategory that can lead to cirrhosis and hepatocellular carcinoma, and thereby to a high mortality rate. Currently, there has been no approved drug to treat NAFLD or NASH. The current study has presented RLA8, a novel and balanced quadruple agonist for hepatic lipid metabolism and inflammation-related peroxisome proliferator-activated receptors (PPARs)- α / γ / δ and G protein-coupled receptor 40 (GPR40), as a NASH drug candidate. The efficacy of RLA8 to treat NASH was evaluated in vivo using two mouse models induced by methionine/choline-deficient diet or by high-fat diet, respectively. RLA8 was shown to improve serum alanine aminotransferase and high-density lipoprotein cholesterol levels, reduce hepatic free fatty acid and triglyceride levels, and alleviate insulin resistance. Cytokine and lipoperoxide analysis revealed that RLA8 could reduce oxidative stress and inflammation. Histochemical and morphologic examination of mouse livers showed that RLA8 could improve pathologic changes such as steatosis, ballooning, collagen fiber, and inflammation. Polymerase chain reaction and Western blot analyses proved that RLA8 could result in PPARs and GPR40 activation, accompanied by upregulation of the 5'AMP-activated protein kinase-acetyl-CoA carboxylase pathway and inhibition of the expression of lipogenic genes and proteins, which provided more insights into its action mechanisms. In summary, RLA8 has significantly better efficacy to improve NASH-induced liver damage such as steatosis, inflammation, and fibrosis, and, consequently, it represents a new and highly promising NASH drug candidate that is worthy of further investigation and development., (Copyright © 2019 by The American Society for Pharmacology and Experimental Therapeutics.)

Super-enhancers (SEs) are enriched with a cluster of mediator binding sites, which are major contributors to cell-type-specific gene expression. Currently, a large quantity of long non-coding RNAs has been found to be transcribed from or to interact with SEs, which constitute super-enhancer associated long non-coding RNAs (SE-lncRNAs). These SE-lncRNAs play essential roles in transcriptional regulation through controlling SEs activity to regulate a broad range of physiological and pathological processes, especially tumorigenesis. However, the pathological functions of SE-lncRNAs in tumorigenesis are still obscure. In this paper, we characterized 5056 SE-lncRNAs and their associated genes by analysing 102 SE data sets. Then, we analysed their expression profiles and prognostic information derived from 19 cancer types to identify cancer-related SE-lncRNAs and to explore their potential functions. In total, 436 significantly differentially expressed SE-lncRNAs and 2035 SE-lncRNAs with high prognostic values were identified. Additionally, 3935 significant correlations between SE-lncRNAs and their regulatory genes were further validated by calculating their correlation coefficients in each cancer type. Finally, the SELER database incorporating the aforementioned data was provided for users to explore their physiological and pathological functions to comprehensively understand the blocks of living systems., (© The Author(s) 2019. Published by Oxford University Press.)

Aim: This study aims to establish a convenient and practical predelivery scoring system for trial of labor after cesarean section (TOLAC)., Methods: The data of 498 patients undergoing TOLAC were retrospectively studied. Indices with statistically significant differences, including cervical score, fetal weight, fetal pelvic index, body mass index and age, were selected. Combined with the presence of vaginal delivery history and indications of the previous cesarean section in these patients, three prenatal forecast scales for vaginal birth after cesarean (VBAC) were established. The receiver operating characteristic curve was drawn, and the best cut-off point was determined. Then, the areas under the curve of the three forecasting methods were compared. The scoring method with the largest area under the curve was considered the best method., Results: The six indications of cesarean section used for the forecasting scale were as follows: cervical score, fetal weight, body mass index, age, presence of vaginal delivery history and the presence of previous obstructive dystocia. The scale that had the largest area under the curve was considered the best forecasting scale., Conclusion: The prenatal forecasting method for TOLAC was preliminarily investigated. It was determined that the scale with six indicators, such as the cervical score, could be used for the prenatal evaluation of TOLAC, providing a predictive basis for the possible success of the trial production for pregnant women. The method and process of VBAC section in our hospital was safe and effective., (© 2018 Japan Society of Obstetrics and Gynecology.)

Objective: To explore the SNP effects of patatin-like phospholipase domain which containing 3 (PNPLA3), transmembrane 6 superfamily member 2 (TM6SF2) gene, environmental effects of smoking, alcohol drinking and interaction between gene-gene, gene-environment and drinking-smoking on hepatitis B virus-associated hepatocellular carcinoma (HBV-HCC). Methods: We collected anticoagulant peripheral blood from patients of HBV-HCC, chronic hepatitis B (CHB), liver cirrhosis (LC) and from healthy controls to detect the single nucleotide polymorphism (SNP) of patatin-like phospholipase domain containing 3 (PNPLA3) gene loci rs738409 and transmembrane 6 superfamily member 2 (TM6SF2) gene loci rs58542926, using the flight mass spectrometry method. The optimal assignment value of gene polymorphisms was defined by using the online SNP stats. Hardy-Weinberg (H-W) balance was tested for SNP. Effects of the genetic and environmental factors to HBV-HCC were analyzed by using the multiple classification logistic regression method. The gene-gene, gene-smoking and alcohol drinking interaction effects were investigated by Fork-Life analysis and binary logistic regression methods. Results: The frequency distribution of CHB group rs738409 loci seemed not in conformity with the H-W balance ( χ (2)=11.980, P <0.005). Two loci frequency distributions in the other groups were all in accordandce with the H-W balance. After adjusting for influences on age and sex and comparing to the healthy group, the rs58542926 mutation appeared as OR =1.659, 95 %CI : 1.026-2.684, P =0.039, in the HBV-HCC group. When comparing to CHB group, the HBV-HCC group presented that drinking as OR =1.680, 95 %CI : 1.121-2.519, P =0.012. When comparing to the LC group, the OR s of drinking and smoking were 1.539 (1.071-2.213) and 1.453 (1.005-2.099) respectively, in the HBV-HCC group. When comparing to the CHB+LC group, interactions between the HBV-HCC group were found rs738409 and rs58542926 on additive model OR =1.548 ( U =1.885, P =0.029) and OR =1.658 ( P =0.024) on logistic regression model while drinking was rs738409 on interaction additive model with OR =1.811( U =1.965, P =0.024). As for drinking and mutation of rs738409, the multiplication model of logistic regression showed no statistically significant differences. Interaction between smoking and drinking appeared as OR =1.756 ( P <0.001) in the logistics regression multiplication model. Conclusions: Factors as mutation of TM6SF2, smoking and drinking all appeared as risk factors for HBV-HCC. Mutations of both PNPLA3 and TM6SF2, together with smoking and drinking all served as risk factors for HBV-HCC. However, the mutation of single PNPLA3 appeared as a protective factor on HBV-HCC.

The purpose of this study was to investigate the occurrence of plasmid-mediated colistin resistance gene mcr-1 in Enterobacteriaceae isolates from companion animals in Guangzhou, China. Enterobacteriaceae isolated from 180 samples collected from cats and dogs were screened for mcr-1 by PCR and sequencing. MCR-1-producing isolates were further characterized by multilocus sequence typing and pulsed-field gel electrophoresis (PFGE). Plasmid characterization was performed by conjugation, replicon typing, S1-PFGE, and Southern blot hybridization. Plasmid pHN6DS2 as a representative IncN1-IncHI2/ST3 plasmid from ST93 E. coli was fully sequenced. pHN6DS2-like plasmids were screened by PCR-mapping and sequencing. The mcr-1 gene was detected in 6.25% (8/128) Escherichia coli isolates, of which, five belonged to E. coli ST93 and had identical PFGE patterns, resistance profiles and resistance genes. mcr-1 genes were located on ∼244.4 kb plasmids ( n = 6), ∼70 kb plasmids, and ∼60 kb plasmids, respectively. Among them, five mcr-1 -carrying plasmids were successfully transferred to recipient by conjugation experiments, and were classified as IncN1-IncHI2/ST3 (∼244.4 kb, n = 4, all obtained from E. coli ST93), and IncI2 (∼70 kb, n = 1), respectively. Plasmid pHN6DS2 contained a typical IncHI2-type backbone, with IncN1 segment (Δ repA- Iterons I- gshB -ΔIS 1294 ) inserted into the multiresistance region, and was similar to other mcr-1 -carrying IncHI2/ST3 plasmids from Enterobacteriaceae isolates of various origins in China. The remaining five mcr-1 -bearing plasmids with sizes of ∼244.4 kb were identified to be pHN6DS2-like plasmids. In conclusion, clonal spread of ST93 E. coli isolates was occurred in companion animals in Guangzhou, China.

To understand the underlying evolution process of F33:A-:B- plasmids among Enterobacteriaceae isolates of various origins in China, the complete sequences of 17 bla CTX-M -harboring F33:A-:B- plasmids obtained from Escherichia coli and Klebsiella pneumoniae isolates from different sources (animals, animal-derived food, and human clinics) in China were determined. F33:A-:B- plasmids shared similar plasmid backbones comprising replication, leading, and conjugative transfer regions and differed by the numbers of repeats in yddA and traD and by the presence of group II intron, except that pHNAH9 lacked a large segment of the leading and transfer regions. The variable regions of F33:A-B- plasmids were distinct and were inserted downstream of the addiction system pemI / pemK , identified as the integration hot spot among F33:A-B- plasmids. The variable region contained resistance genes and mobile elements or contained segments from other types of plasmids, such as IncI1, IncN1, and IncX1. Three plasmids encoding CTX-M-65 were very similar to our previously described pHN7A8 plasmid. Four CTX-M-55-producing plasmids contained multidrug resistance regions related to that of F2:A-B- plasmid pHK23a from Hong Kong. Five plasmids with IncN and/or IncX replication regions and IncI1-backbone fragments had variable regions related to those of pE80 and p42-2. The remaining five plasmids with IncN replicons and an IncI1 segment also possessed closely related variable regions. The diversity in variable regions was presumably associated with rearrangements, insertions, and/or deletions mediated by mobile elements, such as IS 26 and IS 1294 IMPORTANCE Worldwide spread of antibiotic resistance genes among Enterobacteriaceae isolates is of great concern. F33:A-:B- plasmids are important vectors of resistance genes, such as bla CTX-M-55/-65 , bla NDM-1 , fosA3 , and rmtB , among E. coli isolates from various sources in China. We determined and compared the complete sequences of 17 F33:A-:B- plasmids from various sources. These plasmids appear to have evolved from the same ancestor by mobile element-mediated rearrangement, acquisition, and/or loss of resistance modules and similar IncN1, IncI1, and/or IncX1 plasmid backbone segments. Our findings highlight the evolutionary potential of F33:A-:B- plasmids as efficient vectors to capture and diffuse clinically relevant resistance genes., (Copyright © 2018 Wang et al.)

The purpose of this study was to investigate the prevalence and genetic elements of oqxAB among Escherichia coli isolates from animals, retail meat, and humans (patients with infection or colonization) in Guangzhou, China. A total of 1,354 E. coli isolates were screened for oqxAB by PCR. Fifty oqxAB -positive isolates were further characterized by pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), S1-PFGE, genetic environment analysis, plasmid replicon typing, and plasmid sequencing. oqxAB was detected in 172 (33.79%), 60 (17.34%), and 90 (18.07%) E. coli isolates from animal, food, and human, respectively. High clonal diversity was observed among oqxAB -positive isolates. In 21 oqxAB -containing transformants, oqxAB was flanked by two IS 26 elements in the same orientation, formed a composite transposon Tn 6010 in 19 transformants, and was located on plasmids (33.3~500 kb) belonging to IncN1-F33:A-:B- ( n = 3), IncHI2/ST3 ( n = 3), F-:A18:B- ( n = 2), F-:A-:B54 ( n = 2), or others. Additionally, oqxAB was co-located with multiple resistance genes on the same plasmid, such as aac(6 ' )-Ib-cr and/or qnrS , which were identified in two F-:A18:B- plasmids from pigs, and bla CTX-M-55 , rmtB, fosA3 , and floR , which were detected in two N1-F33:A-:B- plasmids from patients. The two IncHI2/ST3 oqxAB -bearing plasmids, pHNLDF400 and pHNYJC8, which were isolated from human patient and chicken meat, respectively, contained a typical IncHI2-type backbone, and were similar to each other with 2-bp difference, and also showed 99% identity to the Salmonella Typhimurium oqxAB -carrying plasmids pHXY0908 (chicken) and pHK0653 (human patient). Horizontal transfer mediated by mobile elements may be the primary mechanism underlying oqxAB spread in E. coli isolates obtained from various sources in Guangzhou, China. The transmission of identical oqxAB -carrying IncHI2 plasmids between food products and humans might pose a serious threat to public health.

Objectives: To investigate the impact of plasmid-borne oqxAB genes on the development of fluoroquinolone resistance, mutations and bacterial fitness in Escherichia coli ., Methods: MICs and mutation prevention concentrations were compared among E. coli strain TOP10 and two corresponding transformants harbouring the OqxAB-encoding plasmids. Mutants were selected by serial passages with the 0.5-fold MIC of ciprofloxacin, and were randomly selected to determine mutations. Bacterial fitness was evaluated by competition assays in vitro and in vivo ., Results: The oqxAB -carrying plasmids contributed to a 4-8-fold increase in the ciprofloxacin MIC and increased the ciprofloxacin mutation prevention concentration by 8-16-fold. The MIC of ciprofloxacin for the two transformants increased faster than that of E. coli TOP10 by serial passaging. Novel mutations in gyrB (A468P or F458V) were first observed. Mutations in gyrA were distributed at codons 87 and 83 in the two transformants, whereas mutation A119E in gyrA dominated in the TOP10 mutants. Although the two oqxAB -bearing plasmids caused a decrease in fitness in vitro , their fitness increased when combined with more than one chromosomal mutation, and clear biological benefits were observed in vivo . The mutations in gyrB were associated with a fitness cost, which could be compensated for by additional mutations. The novel mutation gyrA ΔS83 significantly reduced biological fitness both in vitro and in vivo , and was thus quickly replaced by more beneficial mutations in the population., Conclusions: The possession of plasmid-borne oqxAB may facilitate the evolution of fluoroquinolone resistance, and the fitness cost of OqxAB-encoding plasmids could be compensated by additional chromosomal mutations., (© The Author 2017. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Objective: To investigate the synergistic lethal effect and mechanism of arsenic trioxide (ATO) and aclacinomycin (ACM) on human acute myeloid leukemia cell line KG-1a. Methods: Colony-forming assay was used to detect the proliferation of KG-1a cells treated with different concentration of ATO and ACM. Compusyn software was used to analyze the synergistic effect of ATO and ACM. Flow cytometry and Wright's staining were used to analyze the apoptotic rate of KG-1a cells induced by combined treatment of ATO and ACM. Western blot was used to determine the expression of proteins associated with apoptosis. Results: The cytotoxicity of arsenic trioxide or aclacinomycin alone was in a dose-dependent manner. Flow cytometry analysis showed that the apoptotic rate of KG-1a cells treated with both 0.4 μmol/L ATO and 10 nmol/L ACM was (34.5±3.1)%, significantly higher than (7.6±1.1)% of 0.4 μmol/L ATO treatment or (18.7±2.3) % of 10 nmol/L ACM treatment alone ( P <0.05). The apoptotic rate of KG-1a cells treated with both 1.5 μmol/L ATO and 37.5 nmol/L ACM was (52.5±4.7)%, significantly higher than (19.1±3.2)% of 1.5 μmol/L ATO treatment or (27.7±2.2)% of 37.5 nmol/L ACM treatment alone ( P <0.05). The apoptotic rate of KG-1a cells treated with both 3.0 μmol/L ATO and 75 nmol/L ACM was (61.3±4.5)%, significantly higher than (29.5±2.5)% of 3.0 μmol/L ATO treatment or (28.6±3.4) % of 75 nmol/L ACM treatment alone ( P <0.05). In addition, the result of Wright's staining showed that combined treatment of ATO and ACM induced a more apparent phenotype of apoptosis when compared with single agent treatment. Compusyn software analysis showed that the combination index (CI) value of combined treatment group was less than 1, which indicated the synergistic effect of these two agents. Conclusions: Combined treatment of ATO and ACM shows a synergistic lethal effect on human acute myeloid leukemia cell line KG-1a via activating the apoptotic pathway, which inhibits cell growth and induces apoptosis.

Background: Literature pertaining to prophylactic inguinal nodal treatment for anal adenocarcinoma in China is scarce., Methods: In this retrospective study, we analyzed 126 patients from 1965 to 2015. Among these, 67 patients received surgery only, 18 patients received chemoradiotherapy only, 27 patients received a combination of both, and the remaining 14 patients received palliative treatment., Results: The median follow up period was 30 months. The 1-year, 3-year, and 5-year overall survival rates were 85.8%, 62.5%, and 43.4%, respectively. The 5-year overall survival was 46.9% for patients with negative inguinal lymph nodes and 19.1% for patients with positive inguinal lymph nodes ( p= 0.007). The overall 5-year inguinal node relapse-free survival was 83.0%. The 5-year inguinal node relapse-free survival was 87.5% for stage I, 86.9% for stage II, and 76.5% for stage III cancers. Among those with negative inguinal nodes, the 5-year inguinal node relapse-free survival was 85.7% for negative regional lymph nodes and 75.4% for positive regional lymph nodes ( p= 0.089)., Conclusion: Inguinal lymph node is a high-risk subclinical area. Prophylactic inguinal nodal treatment is necessary for patients with anal adenocarcinoma irrespective of positive or negative inguinal lymph nodes., Competing Interests: Competing Interests: The authors have declared that no competing interest exists.

Water and nitrogen are two of the most important factors for plant growth and development. However, little is known about effects of N on water translocation between connected bamboo ramets. We performed experiment connected Indocalamus decorus ramets in adjacent pots with different soil water contents and three N levels. We determined antioxidase activities, concentration of osmotic adjustment products, O 2 · - , MDA and photosynthetic pigments, and electrolyte leakage rate in paired unit. When N supply to supporting ramets increased, their electrolyte leakage rates and contents of O 2 · - and MDA significantly increased, while antioxidase activities and contents of osmotic adjustment products and photosynthetic pigments in connected dependent ramets increased markedly as their electrolyte leakage rates and contents of O 2 · - and MDA decreased greatly. When N addition to dependent ramets increased, antioxidant enzyme activity and contents of osmotic adjustment products and photosynthetic pigments decreased in both ramets, but electrolyte leakage rates and O 2 · - and MDA contents increased significantly. Therefore, N addition to either supporting or dependent ramets can improve water integration among I. decorus ramets. N addition to supporting ramets promotes water translocation and alleviates water stress of dependent ramets, but N addition to dependent ramets exacerbates drought stress damage to dependent ramets.

Objective: To grope for an ideal immune strategy in grown-ups via comparison of immunological effects under 4 different vaccination schemes. Methods: Study population was selected by stratified random cluster sampling. A total of 4 different vaccination proposals, including Strategy A (3 doses, 10 μg, administrated repeatedly into the unilateral deltoid muscle at 0-1-6 months), Strategy B (2 doses, 20 μg, administrated into the bilateral deltoid muscles simultaneously), Strategy C (3 doses, 10 μg, administrated repeatedly into the unilateral deltoid muscle at 0-1-2 months) and Strategy D (2 doses, 10 μg, administrated to the bilateral deltoid muscles at the same time), were conducted in Liangzhou, Minqin Gulang, and the Tianzhu Tibetan Autonomic county respectively, in Wuwei city, Gansu province. Under 4 different strategies, post-vaccination immunological effectiveness was evaluated when blood samples of participants collected in the eighth months, post-first injection and in the third year, and tested by enzyme-linked immunoassays and electro-chemiluminescence immunoassay. Chi -squared test and Fisher exact test were used to evaluate the immunological differences between the 4 strategies. Wilcoxon' s signed rank test and Kruskal-Waillis H test were conducted to compare the differences of the geometric mean titers (GMTs) of antibody against HBV surface antigen (anti-HBs) titers. Results: A total of 1 621 eligible participants aged 16 to 60 years old, were recruited for the study. Numbers of administration and gender were testified as the presuming factors for influencing immune effectiveness. The vaccination completion rates were 53.97% and 79.82% in Strategy A and C, respectively, and the difference statistically significant ( P <0.05). In the first year, the protective antibody sero-conversion rates (standardization rate) were 89.21%, 54.88%, 92.11%, and 41.63%, in Strategy A, B, C and D, respectively, and the significant statistically differences emerged ( P <0.05) if Strategy B, C and D were compared with Strategy A (as the gold standard). Over a 3-year follow-up period, the levels of GMTs on protective antibody declined from 179.2 IU/L, 51.6 IU/L, 277.1 IU/L and 10.1 IU/L to 61.3 IU/L, 21.2 IU/L, 31.8 IU/L and 6.0 IU/L in Strategy A, B, C and D, respectively, and the differences of declination on GMTs showed statistically significant differences ( P <0.05) when compared within or between the 4 strategies. Conclusion: The 0-1-2 months' prophylactic schedules (Strategy C) seemed superior to the others, in terms of effectively inducing the protective antibody, with shorter duration of vaccination, persisting longer immunity and having higher rate of completive vaccination, so is worth to be recommended as a feasible immune programme for adults, especially for migrants from the rural regions.

Objectve: To investigate the feasibility of establishing xenografted leukemia model by zebrafish, so as to provide the more direct model in vitro and experimental evidence for study of acute myeloid leukemia and screening of the drugs for targeting therapy., Methods: Acute myeloid leukemia cell line KG-1a was labeled with red fluorescent dye-MitoRed, then the labeled cells were injected into the yolk sac of zebrafish embryos. Morphological observation, cell count and histopathological detection were used to analyse the infiltration and metastasis of KG-1a cells in zebrafish., Results: KG1a cells could proliferate and gradually spread to the entire abdominal cavity of the zebrafish after KG-1a cells were injected into the yolk sac during 1-7, the results of cell counting in vitro also proved a significant proliferation of KG-1a cells in zebrafish, suggesting that the implanted leukemia stem cells could survive, proliferate and spread in zebrafish. Further study showed that the implanted cells could be transfered to the liver of zebrafish, these cells displayed the signature of KG-1a cells by hematoxylin-eosin(HE) staining., Conclusions: Human acute myeloid leukemia cells KG1a can survive, proliferate and migrate in zebrafish, suggesting xenografted leukemia model of zebrafish has been successfully established. This model may be benefitcial for the study of acute myeloid leukemia and the screening of the drugs for targeting therapy of acute myeloid leukemia.

Objective: To investigate the value of low-dose multi-slice CT (MSCT) chest scan in the diagnosis of coal workers' pneumoconiosis. Methods: A total of 90 patients with a confirmed diagnosis of coal workers' pneumoconiosis were enrolled, and under the conditions of fixed tube voltage, pitch, and slice thickness, they underwent CT scan with a normal dose (150 mA) and a low dose (30-50 mA). The quality of images obtained from two scans was compared, and the imaging findings, opacity profusion, stage, and radiation doses were also compared. Results: Compared with the normal-dose scan, low-dose scan increased the image noise, and the images obtained from scans with doses of 30, 40, and 50 mA did not show significant reductions in signal-to-noise ratio or contrast-to-noise ratio ( P >0.05). There was no significant difference in the percentage of image quality between low-dose and normal-dose scans ( P >0.05). There were no significant differences in the percentage of various imaging findings, opacity profusion, or percentage of different stages between low-dose (30, 40, and 50 mA) and normal-dose (150 mA) scans ( P >0.05). Conclusion: There are no significant differences between low-dose MSCT chest scan and normal-dose CT in image quality, imaging findings of coal workers' pneumoconiosis, opacity profusion, and stage. Meanwhile, low-dose MSCT chest scan greatly reduces the radiation dose and can be used to assist the diagnosis and follow-up reexamination of coal workers' pneumoconiosis and cover the shortage of high-kilovoltage chest X-ray.

The aggregation behavior and interactions of yak milk protein were investigated after heat treatments. Skim yak milk was heated at temperatures in the range of 65 to 95°C for 10 min. The results showed that the whey proteins in yak milk were denatured after heat treatment, especially at temperatures higher than 85°C. Sodium dodecyl sulfate-PAGE analysis indicated that heat treatment induced milk protein denaturation accompanied with aggregation to a certain extent. When the heating temperature was 75 and 85°C, the aggregation behavior of yak milk proteins was almost completely due to the formation of disulfide bonds, whereas denatured α-lactalbumin and β-lactoglobulin interacted with κ-casein. When yak milk was heated at 85 and 95°C, other noncovalent interactions were found between proteins including hydrophobic interactions. The particle size distributions and microstructures demonstrated that the heat stability of yak milk proteins was significantly lowered by heat treatment. When yak milk was heated at 65 and 75°C, no obvious changes were found in the particle size distribution and microstructures in yak milk. When the temperature was 85 and 95°C, the particle size distribution shifted to larger size trend and aggregates were visible in the heated yak milk., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Mitochondria are closely associated with cell survival, and it is of interest to determine whether apoptosis pathways, which are mediated by mitochondria, are involved in liver regeneration (LR). To identify the mechanisms underlying mitochondria-mediated apoptosis during rat LR, we used the Rat Genome 230 2.0 Array to investigate changes in gene expression. Next, we searched the GO and NCBI databases for genes associated with apoptosis mediated by mitochondria, and QIAGEN and KEGG databases for any related signaling pathways. The expression profile function (Et) was then used to calculate the activity level of known signaling pathways associated with apoptosis. The results revealed the expression of 436 genes associated with apoptosis signaling pathways, among which 152 were confirmed to be primarily related to LR. Overall, 99, 136, 95, and 91 genes were first expressed during the initiation [0.5-4 h after partial hepatectomy (PH)], G0/G1 transition (4-6 h after PH), cell proliferation (6-66 h after PH), and redifferentiation and structural reconstruction (66-144 h after PH) phases, demonstrating that LR-related genes were primarily induced in the initiation phase, and were then expressed across multiple phases. Analysis using the gene synergy formula (Et) showed that caspase-dependent and DNA fragment-related/unrelated pathways induced apoptosis in the early and late periods of LR, and the caspase-independent and DNA fragment-related/unrelated pathways almost in the whole process. Therefore, these results show that several apoptosis pathways regulate LR in rat., Competing Interests: The authors declare no conflict of interest.

The purpose of this study was to identify the optimal local treatment modalities for International Federation of Gynecology and Obstetrics (FIGO) stage I-II small-cell carcinoma of the cervix (SCCC), including cancer-directed surgery (CDS) and/or radiotherapy (RT). The Surveillance Epidemiology and End Results (SEER) database was used to identify SCCC patients from 1988 to 2012, and analyzed using Kaplan-Meier survival and Cox regression proportional hazard methods to determine factors significant for cause-specific survival (CSS) and overall (OS). A total of 208 patients of SCCC were enrolled. The median follow-up time was 31 months. Fifty-eight (27.9%) patients were treated with primary CDS, 88 (42.3%) patients underwent CDS combined with RT, and 62 (29.8%) patients were treated with primary RT. Univariate and multivariate analyses showed that local treatment modalities were independent prognostic factors for CSS and OS. Patients who had undergone CDS had better CSS and OS, compared with patients who had been treated with combined CDS and RT or RT alone. The 5-year CSS and OS of entire group was 49.8% and 46.4%, respectively. The 5-year CSS in the groups of patients receiving CDS, CDS combined with RT, and RT alone were 67.9%, 49.7%, and 32.6%, respectively (P < 0.001). The 5-year OS in patients treated with CDS, CDS combined with RT, and RT alone were 64.9%, 46.2%, and 28.8% (P < 0.001). Primary surgery was associated with improved CSS and OS for FIGO stage I and lymph node negative disease. Primary surgery is the most effective local treatment for FIGO stage I-II SCCC, as adjuvant RT or radical RT does not improve survival compared to radical surgery, especially in patients with FIGO stage I and lymph node negative disease., (© 2016 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.)

Mercury is harmful to the environment, which has gradually become one of the research hotspots. Sediments, as a main repository of pollutants, have an important impact on water quality and the internal organisms, which deserves our research. In this paper, we focused on Hefei landscape water sediment and tried to investigate the status of inorganic mercury and methylmercury pollutions in the sediment. To study the conversion process from inorganic mercury to methylmercury and their enrichment levels and mechanism, we established the ecological chain of "sediment-water-grass-fish" through analog microcosm examination. The results were as follows: from ten water and sediment samples in Hefei landscape water sediment, we found that the contents of inorganic mercury and methylmercury ranged 11.74-13.12 µg · kg⁻¹ and 0.37-2.23 µg · kg⁻¹, respectively. The microcosm examination showed that: with increasing culture time, inorganic mercury in sediments gradually decreased. There was a phenomenon that the content of methylmercury increased at first and then decreased to reach the balance later. Both the inorganic mercury and methylmercury in water change showed an increasing trend. The enrichment contents of inorganic mercury in Egeria densa Planch, and golden mandarin fish (Siniperca scherzeri Steindachner) were low while their enrichment of methylmercury could he great. In addition, we found that both the bioaccumulation ability and the enrichment coefficient of methylmercury in the body of golden mandarin fish were the maximum during the same period.

The research was to approach the growth strategy of rhizome and roots based on the morphology, biomass and nutrient in Phyllostachys rivalis under long-term waterlogged conditions, and provided a theoretical basis for its application for vegetation restoration in wetland and water-level fluctuation belts. The morphological characteristics, physiological and biochemical indexes of annual bamboo rhizome and roots were investigated with an experiment using individually potted P. rivalis which was treated by artificial water-logging for 3, 6, and 12 months. Accordingly the morphological characteristics, biomass allocation, nutrient absorption and balance in rhizome and roots of P. rivalis were analyzed. The results showed that there was no obvious impact of long-term water-logging on the length and diameter of rhizomes, diameter of roots in P. rivalis. The morphological characteristics of rhizome had been less affected generally under water-logging for 3 months. And less rhizomes were submerged, while the growth of roots was inhibited to some extent. Furthermore, with waterlogging time extended, submerged roots and rhizomes grew abundantly, and the roots and rhizomes in soil were promoted. Moreover for ratios of rhizome biomass in soil and water, there were no obvious variations, the same for the root biomass in soil to total biomass. The ratio of root biomass in water to total biomass and the ratio of root biomass in water to root biomass in soil both increased significantly. The results indicated that P. rivalis could adapt to waterlogged conditions gradually through growth regulation and reasonable biomass distribution. However, the activity of rhizome roots in soil decreased and the nutrient absorption was inhibited by long-term water-logging, although it had no effect on stoichiometric ratios of root nutrient in soil. The activity of rhizome root in water increased and the stoichiometric ratios adjusted adaptively to waterlogged conditions, the ratio of N/P increased, while N/K and P/K decreased, which implied that roots in water absorbed oxygen and nutrients could help P. rivalis adapt to long-term waterlogged environment effectively.

Background: The ability of circulating microRNAs (miRNAs) to detect preclinical hepatocellular carcinoma has not yet been reported. We aimed to identify and assess a serum miRNA combination that could detect the presence of clinical and preclinical hepatocellular carcinoma in at-risk patients., Methods: We did a three-stage study that included healthy controls, inactive HBsAg carriers, individuals with chronic hepatitis B, individuals with hepatitis B-induced liver cirrhosis, and patients with diagnosed hepatocellular carcinoma from four hospitals in China. We used array analysis and quantitative PCR to identify 19 candidate serum miRNAs that were increased in six patients with hepatocellular carcinoma compared with eight control patients with chronic hepatitis B. Using a training cohort of patients with hepatocellular carcinoma and controls, we built a serum miRNA classifier to detect hepatocellular carcinoma. We then validated the classifiers' ability in two independent cohorts of patients and controls. We also established the classifiers' ability to predict preclinical hepatocellular carcinoma in a nested case-control study with sera prospectively collected from patients with hepatocellular carcinoma before clinical diagnosis and from matched individuals with hepatitis B who did not develop cancer from the same surveillance programme. We used the sensitivity, specificity, and area under the receiver operating characteristic curve (AUC) to evaluate diagnostic performance, and compared the miRNA classifier with α-fetoprotein at a cutoff of 20 ng/mL (AFP20)., Findings: Between Aug 1, 2009, and Aug 31, 2013, we recruited 257 participants to the training cohort, and 352 and 139 participants to the two independent validation cohorts. In the third validation cohort, 27 patients with hepatocellular carcinoma and 135 matched controls were included in the nested case-control study, which ran from Aug 1, 2009, to Aug 31, 2014. We identified a miRNA classifier (Cmi) containing seven differentially expressed miRNAs (miR-29a, miR-29c, miR-133a, miR-143, miR-145, miR-192, and miR-505) that could detect hepatocellular carcinoma. Cmi showed higher accuracy than AFP20 to distinguish individuals with hepatocellular carcinoma from controls in the validation cohorts, but not in the training cohort (AUC 0·826 [95% CI 0·771-0·880] vs 0·814 [0·756-0·872], p=0·72 in the training cohort; 0·817 [0·769-0·865] vs 0·709 [0·653-0·765], p=0·00076 in validation cohort 1; and 0·884 [0·818-0·951] vs 0·796 [0·706-0·886], p=0·042 for validation cohort 2). In all four cohorts, Cmi had higher sensitivity (range 70·4-85·7%) than did AFP20 (40·7-69·4%) to detect hepatocellular carcinoma at the time of diagnosis, whereas its specificity (80·0-91·1%) was similar to that of AFP20 (84·9-100%). In the nested case-control study, sensitivity of Cmi to detect hepatocellular carcinoma was 29·6% (eight of 27 cases) 12 months before clinical diagnosis, 48·1% (n=13) 9 months before clinical diagnosis, 48·1% (n=13) 6 months before clinical diagnosis, and 55·6% (n=15) 3 months before clinical diagnosis, whereas sensitivity of AFP20 was only 7·4% (n=2), 11·1% (n=3), 18·5% (n=5), and 22·2% (n=6) at the corresponding timepoints (p=0·036, p=0·0030, p=0·021, p=0·012, respectively). Cmi had a larger AUC than did AFP20 to identify small-size (AUC 0·833 [0·782-0·883] vs 0·727 [0·664-0·792], p=0·0018) and early-stage (AUC 0·824 [0·781-0·868] vs 0·754 [0·702-0·806], p=0·015) hepatocellular carcinoma and could also detect α-fetoprotein-negative (AUC 0·825 [0·779-0·871]) hepatocellular carcinoma., Interpretation: Cmi is a potential biomarker for hepatocellular carcinoma, and can identify small-size, early-stage, and α-fetoprotein-negative hepatocellular carcinoma in patients at risk. The miRNA classifier could be valuable to detect preclinical hepatocellular carcinoma, providing patients with a chance of curative resection and longer survival., Funding: National Key Basic Research Program, National Science and Technology Major Project, National Natural Science Foundation of China., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Vascular injury after chronic hypoxia leads to endothelial injury and structural damage to tight junctions (TJs), thereby resulting in a variety of cardiovascular diseases. Thus, attenuating hypoxia-induced damage has great significance for the prevention and treatment of cardiovascular disease. The aim of this study was to investigate whether the endothelial protection conferred by tongxinluo (TXL), a traditional Chinese medicinal compound, is related to its regulation of TJ protein expression. In vivo, we found that TXL could promote hypoxia-induced angiogenesis in lung and liver tissue. In vitro, we found that CoCl2 treatment significantly reduced the expression of the TJ proteins occludin, claudin-1, VE-cadherin, and beta-catenin in cultured human cardiac microvascular endothelial cells. TXL pretreatment abrogated the CoCl2-induced downregulation of these TJ proteins. Conversely, overexpression of Krüppel-like factor 4 (KLF4) inhibited the expression of TJ proteins in human cardiac microvascular endothelial cells, an effect that was reversed by TXL pretreatment. Further experiments showed that TXL could promote endothelial cell proliferation by increasing KLF4 phosphorylation, thereby reversing the effect of KLF4 on the expression of TJ proteins. These findings provide a new molecular mechanism for the TXL-induced increase in TJ protein expression.

Tongxinluo (TXL) is a compound prescription formulated according to the meridian theory of traditional Chinese medicine. It may play an important role in cardiovascular protection by improving endothelial cell function. The aim of present study was to investigate whether endothelial protection with TXL is related to its regulation of tight junction protein expression. Human cardiac microvascular endothelial cells (HCMECs) were cultured and treated with 10(-7)  mol l(-1) angiotensin II (Ang II) and the different doses of TXL; the expression of tight junction proteins occludin, claudin, VE-cadherin and beta-catenin was determined by Western blotting and real-time PCR. Gain-of-function and loss-of-function of Krüppel-like factor 5 (KLF5) were carried out in HCMEC transfected with either KLF5 adenovirus pAd-KLF5 or siRNA specific for KLF5. Angiotensinogen transgenic mice were treated with TXL by oral administration of TXL of 0.75 g kg(-1)  day(-1) , and immunohistochemical staining was performed with antioccludin, anticlaudin, anti-VE-cadherin, antibeta-catenin and anti-KLF5 antibodies. Ang II treatment significantly reduced the expression of tight junction proteins occludin, claudin, VE-cadherin and beta-catenin in cultured HCMECs. TXL pretreatment could abrogate the down-regulation of these tight junction proteins induced by Ang II. Ang II treatment also decreased KLF5 expression at the mRNA and protein levels; TXL pretreatment markedly reversed the inhibitory effect of Ang II on KLF5 expression. Gain-of-function and loss-of-function of KLF5 showed that KLF5 mediated the expression of tight junction proteins in HCMECs. TXL-enhanced expression of the tight junction proteins was mediated by KLF5. In angiotensinogen transgenic mice, TXL also increased the tight junction protein levels by inducing KLF5 expression. Chinese medicine TXL increases tight junction protein levels by inducing KLF5 expression in microvascular endothelial cells., (Copyright © 2015 John Wiley & Sons, Ltd.)

To understand the physiological adaptive mechanism of Phyllostachys violascens to intensive mulching management, the effect of mulching management (CK, 1, 3 and 6 years) on the concentrations and ratios of non-structural carbohydrates (NSC), nitrogen (N) and phosphorus (P) in bamboo foliage, and their stoichiometry was investigated. The results showed the concentrations of NSC and soluble sugar increased, while the starch content and N/P decreased markedly in bamboo stand with 1-year mulching, compared to CK stand, which suggested the N limitation to bamboo growth was strengthened. Foliar soluble sugar content decreased significantly, while the starch content increased dramatically, and the NSC content by per unit mass of N and P reached the maximum in the bamboo stand with 3-year mulching, compared to all other treatments. Foliar NSC and soluble sugar contents decreased significantly, while foliar starch content and N/P increased dramatically in the stand with 6-year mulching, which suggested the P limitation to bamboo growth was strengthened. Foliar NSC content was positively correlated with N and P concentrations in a short-term mulching management stand (≤ 3 years), while showed negative relationship with N/P. The foliar starch content in the stand with 6-year mulching was negatively correlated with N and P contents, while was positively correlated with N/P. The results indicated that short-term mulching management accelerated the accumulation of soluble sugar and decomposition of starch in foliage, thus the growth and activity of Ph. violascens was enhanced greatly. Long-term mulching management promoted the starch accumulation, which led to the transition from N limitation to P limitation for bamboo growth. In summary, long-term (6 years) mulching management caused the decrease of growth and activity of Ph. violascens dramatically, thus enhancing the bamboo stand degradation. The utilization efficiency of N and P reached the highest in the stand with 3-year mulching, which implied 3-year was the best suitable period for intensive mulching management for maintaining bamboo stand quality.

MicroRNAs (miRNAs) play an important role in the regulation of gene expression. Previous studies on miRNA functions mainly focused on their target sites in the 3' untranslated regions (UTRs) of mRNAs. However, increasing evidence has revealed that miRNAs can also induce mRNA degradation and mediate translational repression via complementary interactions with the coding sequence (CDS) and 5'UTR of mRNAs. In this study, we developed a novel database, MtiBase, to facilitate the comprehensive exploration of CDS- and 5'UTR-located miRNA target sites identified from cross-linking immunoprecipitation sequencing (CLIP-Seq) datasets and to uncover their regulatory effects on mRNA stability and translation from expression profile datasets. By integrating 61 Argonaute protein-binding CLIP-Seq datasets and miRNA target sites predicted by five commonly used programs, we identified approximately 4 400 000 CDS-located and 470 000 5'UTR-located miRNA target sites. Moreover, we evaluated the regulatory effects of miRNAs on mRNA stability and translation using the data from 222 gene expression profiles, and 28 ribosome-protected fragment sequencing, and six pulsed stable isotope labeling with amino acids in culture. Finally, the effects of SNPs on the functions of miRNA target sites were systematically evaluated. Our study provides a useful tool for functional studies of miRNAs in regulating physiology and pathology. Database URL: http://mtibase.sysu.edu.cn., (© The Author(s) 2015. Published by Oxford University Press.)

A chelation-directed self-sorting synthesis of a series of cationic heterometallic coordination cages (HCCs) with tunable sizes is described. Two complexation modes were found in the cage-forming process. Metal-anchoring host-guest behavior and size-selective in-cage catalytic activities were found for the HCCs.

Peroxisome proliferator activated receptor γ (PPAR-γ) plays important roles in cell cycle regulation, differentiation and apoptosis. Krüppel-like factor 4 (KLF4) modulates vascular smooth muscle cell (VSMC) phenotype. Both KLF4 and PPAR-γ are involved in VSMC proliferation and differentiation. However, the actual relationship between KLF4 and PPAR-γ in VSMCs is not clear. In this study, we found that PPAR-γ agonist pioglitazone increases KLF4 protein levels but does not influence KLF4 gene transcription. PPAR-γ overexpression increases, while PPAR-γ knockdown reduces KLF4 expression, suggesting that the increase in KLF4 protein levels induced by pioglitazone is PPAR-γ-dependent. Further study showed that pioglitazone enhances KLF4 protein stability through reducing KLF4 ubiquitination. Furthermore, we demonstrated that stabilization of KLF4 by pioglitazone was related to the activation of Akt signaling pathway. Taken together, we revealed that PPAR-γ agonist pioglitazone stabilizes KLF4 protein via activating Akt signaling and reducing KLF4 ubiquitination, providing further insights into PPAR-γ and KLF4 in regulating each other's expression in VSMCs., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Within China's Poyang Lake region, close interactions between wild migratory birds and domestic poultry are common and provide an opportunity for the transmission and subsequent outbreaks of highly pathogenic avian influenza (HPAI) virus. We overlaid a series of ecological factors associated with HPAI to map the risk of HPAI in relation to natural and anthropogenic variables, and we identified two hotspots for potential HPAI outbreaks in the Poyang Lake region as well as three corridors connecting the two hotspot areas. In hotspot I, there is potential for migratory birds to bring new avian influenza (AI) strains that can reassort with existing strains to form new AI viruses. Hotspot II features high-density poultry production where outbreaks of endemic AI viruses are likely. The three communication corridors that link the two hotspots further promote HPAI H5N1 transmission and outbreaks and lead to the persistence of AI viruses in the Poyang Lake region. We speculate that the region's unevenly distributed poultry supply-and-demand system might be a key factor inducing HPAI H5N1 transmission and outbreaks in the Poyang Lake region.

By using open-top chambers (OTCs) to simulate the scenes of elevated CO2 concentrations [500 micromol x mol(-1) (T1) and 700 micromol x mol(-1) (T2)], and taking ambient atmospheric CO2 concentration as the control (CK), this paper studied the effects of elevated CO2 concentration on the lipid peroxidation and anti-oxidation enzyme system in Indocalamus decorus, Pleioblastus kongosanensis, and Sasa glabra leaves. After 103 days treatment, the O2(-)* and MDA contents, relative electron conduction, and soluble sugar content in the three dwarf ornamental bamboo species leaves in T1 had no obvious change, but the activities of anti-oxidation enzymes (SOD, POD, CAT, and APX) changed to a certain extent. In T2, the MDA content and relative electron conduction had no obvious change, but the O2(-)* and soluble sugar contents and the anti-oxidation enzymes activities changed obviously. The adaptation capacity of the three bamboo species to elevated CO2 concentration was in the order of I. decorus > P. kongosanensis > S. glabra.

Background: The distinct expression pattern of tumour-associated antigens (TAAs) might be a critical reason for the inefficacy of immunity-based treatments and heterogeneous postsurgical recovery in patients with solid tumours, including hepatocellular carcinoma (HCC). However, little is known about the clinical value of the coexpression patterns of multiple TAAs., Methods: We determined the expression of multiple TAAs with identified immunogenicity (GPC3, AFP, SSX-2, NY-ESO-1, EpCAM, midkine) and the density of tumour-infiltrating immune cells by immunohistochemistry in a panel of 362 primary HCC patients. We evaluated the association between the TAAs, immune cell infiltration, clinicopathological parameters, and prognosis., Results: Patients who coexpressed more TAAs had better prognosis (P<0.00001, overall survival). The integrated pattern of TAA was associated with good differentiation and small tumour size, and with more CD57(+) natural killer and CD20(+) B-cell infiltration (P<0.05). Multivariate Cox proportional hazards analysis identified the TAA index as an independent prognostic indicator (hazard ratio 0.625; 95% confidence interval 0.467-0.837; P=0.002), and could further predict patient prognosis in collaboration with local immune infiltration., Conclusion: Our results could provide new evidence for the improvement of prognostic molecular signatures in HCC, and a novel rationale for patient enrolment in future immunotherapeutic trials and/or clinical treatments.

Taking pure Oligostachyum lubricum forest as test object, this paper studied the matured and withered leaves carbon (C), nitrogen (N), and phosphorus (P) stoichiometry and N and P resorption patterns of 1-3 years old stands at the densities of 24600-29800 stem hm-2 (D, ), 37500-42600 stem hm-2 (D2 ), 46500 - 52800 stem hm-2 (D3), and 76500 - 85500 stem hm-2 (D4). With increasing stand density, the matured leaves C, N, and P contents and withered leaves C and P contents had an overall decrease, the withered leaves N content decreased after an initial increase, and the matured leaves C content at density )4 decreased dramatically. The leaf C/N and C/P ratio increased with increasing stand density, whereas the leaf N/P ratio increased first but decreased then. At stand densities D3 and D4, the leaf N and P utilization efficiencies were significantly higher than those at D, and D2. With increasing stand density, the leaf N resorption capacity increased after an initial decrease, while the leaf P resorption capacity increased steadily. At stand densities D,-D3, the matured leaves N/P ratio was 16.24-19.37, suggesting that the P limitation occurred, leaf establishment increased, and population increase and expansion enhanced. At density D4, the matured leaves N/P ratio was 13.42-15.74, implying that the N limitation strengthened, leaf withering and defoliation increased, and population increase inhibited. All the results indicated that O. lubricum could regulate its leaf C, N and P contents and stoichiometry and enhance the leaf N and P utilization efficiency and resorption capacity to adapt to the severe competition of environment resources at high stand density. In our experimental condition, 46500-52800 stem hm-2 could be the appropriate stand density for O. lubricum management.

A highly regio-, chemo-, diastereo- and enantioselective organocatalytic [4 + 1] annulation of 2-halo-1,3-dicarbonyl compounds with Morita-Baylis-Hillman adducts catalyzed by commercially available, low cost quinidine for the preparation of synthetically unique and medicinally multi-functionalized isoxazoline N-oxides with three stereogenic centers including adjacent quaternary and tertiary stereocenters has been developed. Notably, the unexpected product ethyl 2-((tert-butyldimethylsilyl)oxy)-2-(5,5-diacetyl-3-((methylsulfonyl)oxy)-4-phenylisoxazolidin-3-yl)acetate (8) bearing a quaternary stereocenter and two tertiary stereocenters was obtained from the undocumented 5,5-diacetyl-3-(2-ethoxy-1-hydroxy-2-oxoethyl)-4-phenyl-4,5-dihydroisoxazole 2-oxide (4ba).