Ancrod, a serine protease purified from the venom of Agkistrodon rhodostoma, is highly specific for fibrinogen. It causes anticoagulation by defibrinogenation and has been used as a therapeutic ...anticoagulant for the treatment of moderate to severe forms of peripheral arterial circulatory disorders in a variety of countries. The DNA of ancrod was amplified by recursive PCR with a yeast bias codon and cloned into the pGEM-T Easy vector. In order to achieve a high level secretion and a full activity expression of ancrod in Pichia pastoris (P. pastoris), the P. pastoris protein disulfide bond isomerase (PpPDI) was co-overexpressed in the strain. The secretion characteristics of ancrod with and without PpPDI were examined. With co-overexpression of PpPDI, the production of recombinant ancrod (rAncrod) was increased to 315mg/L in the culture medium, which is twofold higher than the control strain carrying only the ancrod gene. Through purified by Ni super(2+) affinity chromatography and phenyl Sepharose column, the purity of rAncrod was found to be as high as 95.2%. The fibrinogenolytic and zymographic activities of the rAncrod were determined and found to be similar to that of the native protein. This improved expression system can facilitate further studies and the industrial production of ancrod.
Our previous research showed that neuropilin (Nrp) -1highCD4+CD25+Regulatory T cells (Tregs) exhibited primary negative immunoregulation in sepsis induced immune dysfunction. Tuftsin is the typical ...ligand of Nrp-1. Herein, we investigated the potential therapeutic value and mechanisms of tuftsin in sepsis. Sepsis per se markedly decreased the serum concentration of tuftsin, administration of tuftsin improved the survival rate of septic mice with cecal ligation and puncture (CLP). In vitro study, tuftsin prevented the negative immunoregulation of Nrp-1highCD4+CD25+Tregs, including weakening the expression of forkhead/winged helix transcription factor (Foxp)- 3/cytotoxic T lymphocyte associated antigen (CTLA)-4, inhibiting the secretion of transforming growth factor (TGF)-β, and weakening the immunosuppressive function of Nrp-1highCD4+CD25+Tregs to conventional CD4+CD25-T cells. Tuftsin markedly inhibited the demethylation of Foxp3-Tregs specific demethylated region (TSDR) of Nrp-1highCD4+CD25+Tregs. Tuftsin could represent a new potential therapeutic agentia to improve the outcome of septic mice, and associate with preventing the negative immunoregulation of Tregs via Nrp-1.
Zeno behavior in behavior-based system, which is two or more deterministic behaviors make infinite number of discrete transitions in finite time, can cause the motion speed and rotational velocity of ...the mobile robot change precipitately. Furthermore, the sharp shift of different behaviors will even exacerbate the absolute position errors. This paper present a new collision avoidance algorithm based on hybrid zeno behavior automaton which can eliminate the negative effect of the sharp shift of different behaviors. The core theoretical foundations on designing of the compound zeno behavior in hierarchical hybrid behaviors are presented, including the definitions of different type of behaviors and some theorems, which prove the feasibility of the algorithm. After that, the paper focuses on producing emergency behaviors that lead the mobile robot navigation around the obstacles without sharp shift of different behaviors. Simulation results illustrate the good performance of the control algorithms.
Objective： This study aimed to determine if gastric cardia adenocarcinoma （GCA） risk was associated with the lys （A or ＊2） allele at the rs671 （glu5041ys） polymorphism within the aldehyde ...dehydrogenase 2 （ALDH2） gene in a Chinese Han population. We also aimed to investigate ALDH2 genotypic distributions between subjects from high- and low-incidence areas for both GCA and esophageal squamous cell carcinoma （ESCC）. Methods： We designed a case-control study including 2,686 patients with GCA and 3,675 control subjects from high- and low- incidence areas for both GCA and ESCC in China. TaqMan allele discrimination assay was used to genotype the rs671 polymorphism. χ^2 test and binary logistic regression analysis were used to estimate the odds ratios for the development of GCA, and multivariate ordinal logistic regression was used to analyze ALDH2 genotypic distributions among different groups. Results： Compared with ALDH2＊1/＊1 homozygotes, ALDH2＊1/＊2 and ALDH2＊2/＊2 carriers did not increase the risk for GCA in the Chinese Han population （P〉0.05）. Interestingly, the ratio of homozygous or heterozygous ALDH2 ＊2 carriers in high- incidence areas for both GCA and ESCC was lower than that in low-incidence areas （P〈0.001）. Conclusions： Genotypes of rs671 at ALDH2 may not increase GCA susceptibility in Chinese Han populations. In addition, the ALDH2 genotypic distribution differs between Chinese Han populations from high- and low-incidence areas for both GCA and ESCC. Our findings may shed light on the possible genetic mechanism for the dramatic geographic differences of GCA occurrence in China.
Background： Although that glomerulonephritis is the major cause of end-stage renal disease in developing countries such as China, the increasing prevalence of diabetes has contributed to the changing ...spectrum of predialysis chronic kidney disease. Recent studies have revealed an increased proportion of patients with diabetic kidney disease （DKD） in hemodialysis populations in large cities in China. However, studies regarding the clinical phenotype of DKD in China are extremely limited. The incidence, development, and prognosis of diabetic kidney disease （INDEED） study aims to investigate the incidence, progression, and prognosis of DKD, as well as the associated genetic, behavioral, and environmental factors and biomarkers in patients with DKD in China. Methods： INDEED study is a prospective cohort study based on all participants with diabetes in the Kailuan study, which is a general population-based cohort study in northern China. Altogether, over 10,000 participants with diabetes will be followed biennially. Questionnaires documenting general characteristics, behavioral and environmental factors, and medical history will be administrated. Anthropometric measurements and a series of laboratory tests will be peribrmed in one central laboratory. The DNA, plasma, and urine samples of every participant will be stored in a biobank for future research. Conclusions： INDEED study will provide essential information regarding the clinical phenotype and prognosis of patients with DKD in China and will be valuable to identity factors and biomarkers associated with patients with DKD in China.
A multi-stage dual replica bit-line delay （MDRBD） technique is proposed for reducing access time by suppressing the sense-amplifier enable （SAE） timing variation of low voltage static randomaccess ...memory （SRAM） applications. Compared with the traditional technique, this strategy, using statistical theory, reduces the timing variation by using multi-stage ideas, meanwhile doubling the replica bit-fine （RBL） capacitance and discharge path simultaneously in each stage. At a supply voltage of 0.6 V, the simulation results show that the standard deviations of the SAE timing and cycle time with the proposed technique are 69.2% and 47.2%, respectively, smaller than that with a conventional RBL delay technique in TSMC 65 nm CMOS technology （Taiwan Semiconductor Manufacturing Company, Taiwan）.
Ethylene plays essential roles during adaptive responses to water-saturating environments in rice, but knowledge of its signaling mechanism remains limited. Here, through an analysis of a rice ...ethylene-response mutant mhz1, we show that MHZ1 positively modulates root ethylene responses. MHZ1 encodes the rice histidine kinase OsHK1. MHZ1/OsHK1 is autophosphorylated at a conserved histidine residue and can transfer the phosphoryl signal to the response regulator OsRR21 via the phosphotransfer proteins OsAHP1/2. This phosphorelay pathway is required for root ethylene responses. Ethylene receptor OsERS2, via its GAF domain, physically interacts with MHZ1/OsHK1 and inhibits its kinase activity. Genetic analyses suggest that MHZ1/OsHK1 acts at the level of ethylene perception and works together with the OsEIN2-mediated pathway to regulate root growth. Our results suggest that MHZ1/OsHK1 mediates the ethylene response partially independently of OsEIN2, and is directly inhibited by ethylene receptors, thus revealing mechanistic details of ethylene signaling for root growth regulation.
Summary
Soybean (Glycine max) is one of the most important oilseed crops. However, the regulatory mechanism that governs the process of oil accumulation in soybean remains poorly understood.
In this ...study, GmZF392, a tandem CCCH zinc finger (TZF) protein which was identified in our previous RNA‐seq analysis of seed‐preferred transcription factors, was found to function as a positive regulator of lipid production.
GmZF392 promotes seed oil accumulation in both transgenic Arabidopsis and stable transgenic soybean plants by binding to a bipartite cis‐element, containing TG‐ and TA‐rich sequences, in promoter regions, activating the expression of genes in the lipid biosynthesis pathway. GmZF392 physically interacts with GmZF351, our previously identified transcriptional regulator of lipid biosynthesis, to synergistically promote downstream gene expression. Both GmZF392 and GmZF351 are further upregulated by GmNFYA, another transcription factor involved in lipid biosynthesis, directly (in the former case) and indirectly (in the latter case). Promoter sequence diversity analysis showed that the GmZF392 promoter may have been selected at the origin of the Glycine genus and further mildly selected during domestication from wild soybeans to cultivated soybeans.
Our study reveals a regulatory module containing three transcription factors in the lipid biosynthesis pathway, and manipulation of the module may improve oil production in soybean and other oilseed crops.
Soybean is an important crop worldwide, but its production is severely affected by salt stress. Understanding the regulatory mechanism of salt response is crucial for improving the salt tolerance of ...soybean. Here, we reveal a role for nuclear factor Y subunit GmNFYA in salt tolerance of soybean likely through the regulation of histone acetylation. GmNFYA is induced by salt stress. Overexpression of GmNFYA significantly enhances salt tolerance in stable transgenic soybean plants by inducing salt‐responsive genes. Analysis in soybean plants with transgenic hairy roots also supports the conclusion. GmNFYA interacts with GmFVE, which functions with putative histone deacetylase GmHDA13 in a complex for transcriptional repression possibly by reducing H3K9 acetylation at target loci. Under salt stress, GmNFYA likely accumulates and competes with GmHDA13 for interaction with GmFVE, leading to the derepression and maintenance of histone acetylation for activation of salt‐responsive genes and finally conferring salt tolerance in soybean plants. In addition, a haplotype I GmNFYA promoter is identified with the highest self‐activated promoter activity and may be selected during future breeding for salt‐tolerant cultivars. Our study uncovers the epigenetic regulatory mechanism of GmNFYA in salt‐stress response, and all the factors/elements identified may be potential targets for genetic manipulation of salt tolerance in soybean and other crops.
![Co-overexpression of [Bold]...](http://api.summon.serialssolutions.com/2.0.0/image/issn/XR4PS5YY4K/0273-2289/small "Co-overexpression of [Bold][Italic]Pp[/Bold]DI Enhances Secretion of Ancrod in [Bold][Italic]Pichia pastoris[/Bold]")
PDF
