Generation of Hepatitis B virus PreS2-S antigen in Hansenula polymorpha

Despite the long availability of a traditional prophylactic vaccine containing the HBV surface antigen(HBsA g) and aluminum adjuvant, nearly 10% of the population remains unable to generate an effective immune response. Previous studies have indicated that hepatitis B virus(HBV) PreS 2-S is abundant in T/B cell epitopes, which induces a stronger immune response than HBsA g, particularly in terms of cytotoxic T lymphocyte(CTL) reaction. In the current study, the HBV PreS 2-S gene encoding an extra26 amino acids(PreS 2 C-terminus) located at the N-terminus of HBsA g was cloned into the pV CH1300 expression vector. Pre S2-S expressed in the methylotrophic yeast, Hansenula polymorpha, was produced at a yield of up to 250 mg/L. Subsequent purification steps involved hydrophobic adsorption to colloidal silica, ion-exchange chromatography and density ultracentrifugation. The final product was obtained with a total yield of ~15% and purity of ~99%. In keeping with previous studies, ~22 nm viruslike particles were detected using electron microscopy. The generated PreS 2-S antigen will be further studied for efficacy and safty in animals.

Effects of soil fauna on leaf litter decomposition under different land uses in eastern coast of China

Soil fauna decompose litter, whereas land use changes may significantly alter the composition and structure of soil fauna assemblages. However, little is known of the effects of land-use on the contribution of soil fauna to litter decomposition. We studied the impacts of soil fauna on the decomposition of litter from poplar trees under three different land uses （i.e. poplar-crop integrated system, poplar plantation, and cropland）, from December 2013 to December 2014, in a coastal area of Northern Jiangsu Province. We collected litter samples in litterbags with three mesh sizes （5, 1 and 0. 01 mm, respectively） to quantify the contribution of various soil fauna to the decomposition of poplar leaf litter. Litter decomposition rates differed significantly by land use and were highest in the cropland, intermediate in the poplar-crop integrated system, and lowest in the poplar plantation. Soil fauna in the poplar-crop integrated system was characterized by the highest numbers of taxa and individuals, and highest Margalef＇s diversity, which suggested that agro-forestry ecosystems may support a greater quantity, distribution, and biodiversity of soil fauna than can single-species agriculture or plantation forestry. The individuals and groups of soil fauna in the macro-mesh litterbags were higher than in the meso-mesh litterbags underthe same land use types. The average contribution rate of meso- and micro-fauna to litter decomposition was 18.46%, which was higher than the contribution rate of macro-fauna （3.31%）. The percentage of remaining litter mass was inversely related to the density of the soil fauna （P 〈 0.05） in poplar plantations; however, was unrelated in the poplar-crop integrated system and cropland. This may have been the result of anthropogenic interference in poplar-crop integrated systems and croplands. Our study suggested that when land-use change alters vegetation types, it can affect species composition and the structure of soil fauna assemblages, which, in turn, affects litter decomposition.

Integrins as attractive targets for cancer therapeutics

Integrins are transmembrane receptors that have been implicated in the biology of various human physiological and pathological processes.These molecules facilitate cell-extracellular matrix and cell-cell interactions,and they have been implicated in fibrosis,inflammation,thrombosis,and tumor metastasis.The role of integrins in tumor progression makes them promising targets for cancer treatment,and certain integrin antagonists,such as antibodies and synthetic peptides,have been effectively utilized in the clinic for cancer therapy.Here,we discuss the evidence and knowledge on the contribution of integrins to cancer biology.Furthermore,we summarize the clinical attempts targeting this family in anti-cancer therapy development.

Trends in insulin resistance:insights into mechanisms and therapeutic strategy

The centenary of insulin discovery represents an important opportunity to transform diabetes from a fatal diagnosis into a medically manageable chronic condition.Insulin is a key peptide hormone and mediates the systemic glucose metabolism in different tissues.Insulin resistance(IR)is a disordered biological response for insulin stimulation through the disruption of different molecular pathways in target tissues.Acquired conditions and genetic factors have been implicated in IR.Recent genetic and biochemical studies suggest that the dysregulated metabolic mediators released by adipose tissue including adipokines,cytokines,chemokines,excess lipids and toxic lipid metabolites promote IR in other tissues.IR is associated with several groups of abnormal syndromes that include obesity,diabetes,metabolic dysfunction-associated fatty liver disease(MAFLD),cardiovascular disease,polycystic ovary syndrome(PCOS),and other abnormalities.Although no medication is specifically approved to treat IR,we summarized the lifestyle changes and pharmacological medications that have been used as efficient intervention to improve insulin sensitivity.Ultimately,the systematic discussion of complex mechanism will help to identify potential new targets and treat the closely associated metabolic syndrome of IR.

Optimized functional and structural design of dual-target LMRAP,a bifunctional fusion protein with a 25-amino-acid antitumor peptide and GnRH Fc fragment

To develop fusion protein of a GnRH Fc fragment and the integrin targeting AP25 antitumor peptide for GnRH receptor-expressing cancer therapy.The LMRAP fusion protein was constructed.A transwell invasion assay was performed.The gene mRNA and protein levels of GnRHR-I,α5β1,andαvβ3 in different cancer cell lines were assessed.Cell proliferation was measured using a cell counting kit-8.An antagonist assay was performed on GnRH receptors.Anti-tumor activity was evaluated with a mouse xenograft tumor model.Immunohistochemistry(IHC)was applied to detect CD31 and CD34 expressions.Pharmacokinetic characteristics were determined with an indirect competition ELISA.The developed bifunctional fusion protein LMRAP not only inhibited HUVEC invasion,but also inhibited proliferation of GnRHR-I,α5β1,and high expression cancer cells.The IC50 for LMRAP in the GnRH receptor was 6.235 x 10-4 mol/L.LMRAP significantly inhibited human prostate cancer cell line22 RV1 proliferation in vivo and in vitro.LMRAP significantly inhibited CD31 and CD34 expressions.The elimination half-life of the fusion protein LMRAP was 33 h in rats.The fusion protein made of a GnRH Fc fragment and the integrin targeting AP25 peptide retained the bifunctional biological activity of GnRHR blocking,angiogenesis inhibition,prolonged half-life and good tolerance.

Natural killer cell-derived exosomal miR-1249-3p attenuates insulin resistance and inflammation in mouse models of type 2 diabetes

Natural killer(NK)cells have been suggested to be associated with type 2 diabetes by regulating systemic inflammation.However,the mechanism by which NK cells regulate insulin sensitivity remains unknown.This study shows that NK-derived exosomes from lean mice attenuate obesity-induced insulin resistance and inflammation in mice of type 2 diabetes.Moreover,lean NK-derived exosomes enhance insulin sensitivity and relieve inflammation in adipocytes and hepatocytes.MiR-1249-3p,which is significantly upregulated in lean NK-derived exosomes,can be transferred from NK cells to adipocytes and hepatocytes via exosomes.NKderived exosomal miR-1249-3p dramatically induces cellular insulin sensitivity and relieves inflammation.Mechanistically,exosomal miR-1249-3p directly targets SKOR1 to regulate the formation of ternary complex SMAD6/MYD88/SMURF1,which mediates glucose homeostasis by suppressing the TLR4/NF-κB signaling pathway.This study reveals an emerging role for NK-derived exosomal miR1249-3p in remission of insulin resistance,and provides a series of potential therapeutic targets in type 2 diabetes.

Distribution of micropeptide-coding sORFs in transcripts

Small open reading frames(sORFs) are distributed over a wide variety of transcripts. sORFs encoding functional peptides have been identified in various configurations within apparently long noncoding RNAs. Many translated sORFs have been identified across mRNAs, including 5’-upstream, coding domain, and 3’-downstream. sORFs have also been found in circular RNAs, pri-miRNAs, and ribosomal RNAs. Here, we present an overview of the wide distribution of the sORFs in transcripts and their functional roles in organisms.