Circulating histones are major mediators of systemic inflammation and cellular injury in patients with acute liver failure

Acute liver failure (ALF) is a life-threatening systemic disorder. Here we investigated the impact of circulating histones, recently identified inflammatory mediators, on systemic inflammation and liver injury in murine models and patients with ALF. We analyzed histone levels in blood samples from 62 patients with ALF, 60 patients with chronic liver disease, and 30 healthy volunteers. We incubated patients'' sera with human L02 hepatocytes and monocytic U937 cells to assess cellular damage and cytokine production. d-galactosamine plus lipopolysaccharide (GalN/LPS), concanavalin A (ConA), and acetaminophen (APAP) were given to C57BL/6N mice to induce liver injury, respectively, and the pathogenic role of circulating histones was studied. Besides, the protective effect of nonanticoagulant heparin, which can bind histones, was evaluated with in vivo and ex vivo investigations. We observed that circulating histones were significantly increased in patients with ALF, and correlated with disease severity and mortality. Significant systemic inflammation was also pronounced in ALF patients, which were associated with histone levels. ALF patients'' sera induced significant L02 cell death and stimulated U937 cells to produce cytokines, which were abrogated by nonanticoagulant heparin. Furthermore, circulating histones were all released remarkably in GalN/LPS, ConA, and APAP-treated mice, and associated with high levels of inflammatory cytokines. Heparin reduced systemic inflammation and liver damage in mice, suggesting that it could interfere with histone-associated liver injury. Collectively, these findings demonstrate that circulating histones are critical mediators of systemic inflammation and cellular damage in ALF, which may be potentially translatable for clinical use.Acute liver failure (ALF) is a complex condition wherein rapid-onset liver insult results in coagulopathy, hepatic encephalopathy (HE), and even multiple organ failure and death in a patient without previously recognized liver disease.^{1, 2} The causes of ALF vary greatly by geographical region. In the Western countries, acetaminophen (APAP) constitutes nearly 60–70% of the cases, whereas the primary cause of ALF in China is viral hepatitis B, which accounts for 70–80% of the cases.^{3, 4} Despite recent therapeutic advances, ALF remains a serious clinical condition associated with a high mortality rate. The underlying mechanisms of ALF are multifactorial and incompletely understood. Emerging evidence suggests that, regardless of various etiologies of ALF, its acute onset is generally associated with significant and uncontrolled activation of systemic inflammation, which may consequently lead to multiple organ dysfunction and a poorer prognosis in ALF.^{5, 6} It has been accepted that systemic inflammation may have a crucial role in the initiation and progression of ALF, but the key factors or mechanisms that are responsible for its activation in ALF are largely unclear. Therefore, identification of key mediators that may modulate ALF-associated inflammation is highly desirable.Extracellular histones in the circulation are recently identified as the pivotal mediators in systemic inflammatory diseases.^{7, 8, 9, 10} It reveals that circulating histones have numerous toxic effects including direct cytotoxicity, induction of vascular permeability, coagulation activation, platelet aggregation, and cytokine production,^{11, 12, 13} all of which are possible mechanisms related to the development of inflammatory organ injuries. Furthermore, histone-targeted therapy has promising potentials for the treatment of various inflammatory injuries. We, therefore, investigated circulating histone levels in patients with ALF, with a hypothesis that histones present in the circulation of ALF patients could serve as inflammatory mediators mediating cellular damage and interfering with disease progression and mortality. We further examined whether histone-mediated toxicity could be neutralized by nonanticoagulant heparin, which can bind histones,¹⁴ through in vivo and ex vivo translational studies, with an aim to providing an interventional strategy for ALF in clinical practice.     

Two new and one newly recorded species of Gracillariidae from China (Lepidoptera)

The paper presents four Chinese species belonging to the genera Metriochroa Busck, Eumetriochroa Kumata, and Gibbovalva Kumata & Kuroko (Lepidoptera, Gracillariidae), including two new species: Metriochroa alboannulata Bai, sp. n. and Gibbovalva clavata Bai, sp. n. Eumetriochroa hiranoi Kumata, 1998, is newly recorded from China. Photographs of adults and figures of the genital structures are provided, along with keys to the Chinese species of Metriochroa, Eumetriochroa, and Gibbovalva.     

A Novel Strategy for the Preparation of Codon-Optimized Truncated Ulp1 and its Simplified Application to Cleavage the SUMO Fusion Protein

Ubiquitin-like protease 1 (Ulp1) of Saccharomyces cerevisiae emerges as a fundamental tool to obtain the natural N-terminal target protein by cleavage of the small ubiquitin-related modifier (SUMO) fusion protein. However, the costly commercial Ulp1 and its complicated procedures limit its application in the preparation of the target protein with natural N-terminal sequence. Here, we describe the preparation of bioactive codon-optimized recombinant truncated Ulp1 (Leu403-Lys621) (rtUlp1) of S. cerevisiae in Escherichia coli using only one-step with Ni–NTA affinity chromatograph, and the application of rtUlp1 to cleave the SUMO fusion protein by simply mixing the purified rtUlp1, SUMO fusion protein and DL-Dithiothreitol in Tris–HCl buffer. The optimal expression level of non-fusion protein rtUlp1 accounts for approximately 50 % of the total cellular protein and 36 % of the soluble form by addition of isopropyl β-D-l-thiogalactopyranoside at a final concentration of 0.4 mM at 18 °C for 20 h. The purification of target protein rtUlp1 was conducted by Ni–NTA affinity chromatography. The final yield of rtUlp1 was 45 mg/l in flask fermentation with a purity up to 95 %. Furthermore, the high purity of rtUlp1 could effectively cleave the SUMO-tTβRII fusion protein (SUMO gene fused to truncated transforming growth factor-beta receptor type II gene) with the above simplified approach, and the specific activity of the rtUlp1 reached up to 2.8 × 10⁴ U/mg, which is comparable to the commercial Ulp1. The preparation and application strategy of the rtUlp1 with commonly available laboratory resources in this study will be convenient to the cleavage of the SUMO fusion protein to obtain the natural N-terminal target protein, which can be implemented in difficult-to-express protein functional analysis.     

籽用西瓜种质资源SSR分析及初级核心种质库构建

采用SSR分子标记技术对50份不同来源的籽用西瓜材料进行遗传多样性研究,并基于SSR分子标记聚类分析采用最小距离逐步抽样法构建籽瓜初级核心种质库。研究表明：（1）从106对SSR随机引物中筛选出扩增条带清晰、多态性高的31对引物,共得到138条清晰可辨条带,其中多态性条带115条,占83.33%,平均Shannon’s信息指数(I）为0.419 3,平均Nei’s多样指数(h）为0.272 4,平均有效等位基因数(N_e）为1.458 0,平均等位基因数(N_a）为1.981 4,说明50份籽瓜种质具有丰富的遗传多样性。（2）用NTsys2.10e软件对种质资源聚类分析表明,种质间的遗传相似系数介于0.57~0.91之间,其中来源地相同的个别种质间遗传相似系数却很小,而来源地不同的个别种质间的遗传相似系数却很高。（3）采用最小距离逐步抽样法,按70%、60%、50%、40%、30%、20%、10%的比例抽样后,各个核心子集遗传多样性指数总体上变化不明显,但各抽样比例相比,以抽样比例20%获得10份初选种质的 N_e、 h和 I达到最高值,说明抽样20%构建的初级核心种质对原始种质具有很好的代表性。     

Genetic variants in the vitamin D pathway genes VDBP and RXRA modulate cutaneous melanoma disease‐specific survival

Single nucleotide polymorphisms (SNPs) in the vitamin D pathway genes have been implicated in cutaneous melanoma (CM) risk, but their role in CM disease‐specific survival (DSS) remains obscure. We comprehensively analyzed the prognostic roles of 2669 common SNPs in the vitamin D pathway genes using data from a published genome‐wide association study (GWAS) at The University of Texas M.D. Anderson Cancer Center (MDACC) and then validated the SNPs of interest in another GWAS from the Nurses’ Health Study and Health Professionals Follow‐up Study. Among the 2669 SNPs, 203 were significantly associated with DSS in MDACC dataset (P < 0.05 and false‐positive report probability < 0.2), of which 18 were the tag SNPs. In the replication, two of these 18 SNPs showed nominal significance: the VDBP rs12512631 T > C was associated with a better DSS [combined hazards ratio (HR) = 0.66]; and the same for RXRA rs7850212 C > A (combined HR = 0.38), which were further confirmed by the Fine and Gray competing‐risks regression model. Further bioinformatics analyses indicated that these loci may modulate corresponding gene methylation status.     

Should China subsidize cofiring to meet its 2020 bioenergy target? A spatio‐techno‐economic analysis

China has developed ambitious bioenergy installation targets as part of its broader goals to increase its renewable energy‐generating capacity and decarbonize its economy. A key target feedstock for bioenergy is the 800 million tonnes of agricultural residues that China produces each year. At present, the main financial incentive to support bioenergy generation from agricultural residues is a feed‐in‐tariff provided for bioenergy that is produced by units that take 80% or more of their feedstock energy from biomass. Although this policy has catalysed the construction of many bioenergy units, there are reports that these projects are experiencing serious financial and technical problems, leading to low operational efficiency and even closure. An alternative option for China's agricultural residues is cofiring with coal in existing power stations. However, this is currently unprofitable for power station operators, as cofiring is not eligible for financial assistance through the bioenergy feed‐in‐tariff. In the light of China's ambitious target to install 30GW of bioenergy generation capacity by 2020, this study investigates the extent to which extension of the bioenergy feed‐in‐tariff to include cofiring could contribute towards this goal. The results suggest that 39% of China's straw energy resources are located within 50 km of a power station. Assuming cofiring ratios of up to 10% coal energy replacement, an annual 89–117TWh of electricity could be generated by cofiring agricultural residues collected within 50 km radii of power stations. If China extends its bioenergy subsidies to include cofiring, an annual 62–92TWh can be produced at an internal rate of return of 8% or more. This equates to 42–62% of the bioenergy generation that China might expect if it met its 2020 target of installing 30GW of bioenergy capacity. Overall, this indicates a strong case for the Chinese government to extend its existing bioenergy feed‐in‐tariff to include cofiring at low energy replacement ratios.