Facile fabrication of branched-chain carbohydrate chips for studying carbohydrate-protein interactions by QCM biosensor

A novel approach for fabricating branched-chain （BC） carbohydrate chips to study carbohydrate-protein interactions using Quartz Crystal Microbalance （QCM） biosensor was developed. This approach utilizes functional alkynyl-branch molecule modified chip surfaces, which through the terminal alkynyl group for covalent linking of unprotected azide-carbohydrates. The unprotected azide-carbohydrates were syhthesized in one-step using 2-azido-l,B-dimethylqmidazolinium as catalyst, avoiding complex chemical modifications. Additionally, the branch surface modified with the carbohydrates not only supplies more specific binding site but also reveals significant cluster effect. To exemplify the sugar cluster effect on BC carbohydrate chips, BC Galactose and Mannose chips prepared in this work were used to determine carbohydrate-lectin interactions using QCM biosensor. The results clearly showed that BC chip significantly improves the detection sensitivity compared with the straight-chain （SC） chip. More importantly, the BC galactose chip sensitivity was enhanced 40% compared with the SC galactose chip.

Romidepsin(FK228) improves the survival of allogeneic skin grafts through downregulating the production of donor-specific antibody via suppressing the IRE1α-XBP1 pathway

Antibody-mediated rejection(AMR)is one of the major causes of graft loss after transplantation.Recently,the regulation of B cell differentiation and the prevention of donor-specific antibody(DSA)production have gained increased attention in transplant research.Herein,we established a secondary allogeneic in vivo skin transplant model to study the effects of romidepsin(FK228)on DSA.The survival of grafted skins was monitored daily.The serum levels of DSA and the number of relevant immunocytes in the recipient spleens were evaluated by flow cytometry.Then,we isolated and purified B cells from B6mouse spleens in vitro by magnetic bead sorting.The B cells were cultured with interleukin-4(IL-4)and anti-clusters of differentiation 40(CD40)antibody with or without FK228 treatment.The immunoglobulin G1(IgG1)and IgM levels in the supernatant were evaluated by enzyme-linked immunosorbent assay(ELISA).Quantitative reverse transcription-polymerase chain reaction(RT-q PCR)and western blotting were conducted to determine the corresponding levels of messenger RNA(mRNA)and protein expression in cultured cells and the recipient spleens.The results showed that FK228 significantly improved the survival of allogeneic skin grafts.Moreover,FK228 inhibited DSA production in the serum along with the suppression of histone deacetylase 1(HADC1)and HDAC2 and the upregulation of the acetylation of histones H2A and H3.It also inhibited the differentiation of B cells to plasma cells,decreased the transcription of positive regulatory domain-containing 1(Prdm1)and X-box-binding protein 1(Xbp1),and decreased the expression of phosphorylated inositol-requiring enzyme 1α(p-IRE1α),XBP1,and B lymphocyte-induced maturation protein-1(Blimp-1).In conclusion,FK228 could decrease the production of antibodies by B cells via inhibition of the IRE1α-XBP1 signaling pathway.Thus,FK228 is considered as a promising therapeutic agent for the clinical treatment of AMR.

Design and synthesis of novel α-aminoamides derivatives as Nav1.7 inhibitors for antinociception

Three novel series of α-aminoamides derivatives were designed and synthesized based on ralfinamide,and their Nav1.7 inhibitory activities were evaluated using manual patch clamp electrophysiology. Active compounds inhibited Nav1.7 with half maximal inhibitory concentration(IC_(50)) values ranging from2.9 μmol/L to 21.4 μmol/L. Among them, the most potent compound 19h exhibited about 12-fold potency better than ralfinamide. The investigation of their structure-activity relationship gives a strategy to improve the Nav1.7 inhibition of ralfinamide analogues. Compound 19h was efficacious in antinociception in the mouse spared nerve injury(SNI) model of neuropathic pain without causing sedation in the open field test.