RESUMO
Vanin-1 is an amidohydrolase that catalyses the conversion of pantetheine into the amino-thiol cysteamine and pantothenic acid (coenzyme A precursor), which plays a vital role in multiple physiological and pathological processes. In this study, an enzyme-activated near-infrared (NIR) fluorescent probe (DDAV) has been constructed for sensitively detecting Vanin-1 activity in complicated biosamples on the basis of its catalytic characteristics. DDAV exhibited a high selectivity and sensitivity toward Vanin-1 and was successfully applied to the early diagnosis of kidney injury in cisplatin-induced kidney injury model. In addition, DDAV could serve as a visual tool for in situ imaging endogenous Vanin-1 in vivo. More importantly, Enterococcus faecalis 20247 which possessed high expression of Vanin-1 was screened out from intestinal bacteria using DDAV, provided useful guidance for the rational use of NSAIDs in clinic. Finally, oleuropein as a potent natural inhibitor for Vanin-1 was discovered from herbal medicines library using a high-throughput screening method using DDAV, which held great promise for clinical therapy of inflammatory bowel disease.
RESUMO
The glycoproteins in the biological sample are low abundance and are susceptible to be inhibited and interfered by other non-glycoproteins.An enrichment step is usually required before the glycoprotein analysis, but the operation steps of conventional solid-phase-based glycoprotein enrichment methods are difficult to be compatible with the most classical enzyme-linked immune sorbent assay (ELISA) technique.In this study, a novel water-soluble dendrimer based boronic acid capture (DBC) material was developed using PAMAM 4.0 as the carrier and boronic acid as the affinity group.The method was applied to the detection of glycoproteins in human liver microsomes using ELISA.In this study, the DBC enrichment conditions were optimized by model glycoprotein, and then its sensitivity and anti-interference ability were investigated.This method was applied to the enrichment of glycoproteinsin human liver microsomal.The results showed that the enrichment selectivity of DBC for glycoprotein could be up to 100000 folds, and the enrichment signal of glycoprotein could be increased by 100 times.Therefore, the ELISA method using DBC as a novel enrichment material for glycoprotein had high sensitivity and selectivity in detection of biological samples with only one simple incubation step, which was useful for glycoproteins researches.
RESUMO
OBJECTIVE:To investigate the neuropotective effect and anticoagulation effect of total Saponins of Radix Liriopes on focal cerebral ischemia in rats.METHODS:Chemical reagent Fecl3 was locally applied on the injured vessels to establish middle cerebral artery thrombosis model,and the effects of total Saponins of Radix Liriopes on rats' behavioral disturbance,brain infarct size,the histopathological changes in brain and the expression rate of nNOS immunoreactive positive neurons were measured,and the bleeding time and coagulation time were also detected with glass tube method and tail transection method.RESULTS:Due to the use of total Saponin of Radix Liriopes(10 and 40mg? kg-1),the brain infarct size was significantly decreased,the behavioral disturbance were improved and the expression rate of the nNOS immunoreactive positive neurons in rats were decreased.At doses of 20 and 60mg? kg-1,total Saponin of Radix Liriopes significantly prolonged the coagulation time and bleeding time.CONCLUSION:Total Saponin of Radix Liriopes has nuroprotective effect on focal cerebral ischemia induced by middle cerebral artery thrombosis and significant anticoagulation effect.