ABSTRACT
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.
ABSTRACT
Imipenem is a carbapenem antibiotic. However, Imipenem could not be marketed owing to its instability and nephrotoxicity until cilastatin, an inhibitor of renal dehydropeptidase-I (DHP-I), was developed. In present study, the potential roles of renal organic anion transporters (OATs) in alleviating the nephrotoxicity of imipenem by cilastatin were investigated and in rabbits. Our results indicated that imipenem and cilastatin were substrates of hOAT1 and hOAT3. Cilastatin inhibited hOAT1/3-mediated transport of imipenem with IC values comparable to the clinical concentration, suggesting the potential to cause a clinical drug-drug interaction (DDI). Moreover, imipenem exhibited hOAT1/3-dependent cytotoxicity, which was alleviated by cilastatin and probenecid. Furthermore, cilastatin and probenecid ameliorated imipenem-induced rabbit acute kidney injury, and reduced the renal secretion of imipenem. Cilastatin and probenecid inhibited intracellular accumulation of imipenem and sequentially decreased the nephrocyte toxicity in rabbit primary proximal tubule cells. Renal OATs, besides DHP-I, was also the target of interaction between imipenem and cilastatin, and contributed to the nephrotoxicity of imipenem. This therefore gives in part the explanation about the mechanism by which cilastatin protected against imipenem-induced nephrotoxicity. Thus, OATs can potentially be used as a therapeutic target to avoid the renal adverse reaction of imipenem in clinic.
ABSTRACT
In order to clarify regions of production and to discriminate processing methods, quantitative and qualitative analyses for saccharides and terpenes in 35 batches of Alismatis Rhizoma were performed. Methodologies included HPLC-PDA, HPLC-VWD and UHPLC-MS , combined with principal component analysis (PCA) and partial least squares regression techniques (PLSR). The inhibitory effects of triterpenes and Alismatis Rhizoma extracts on lipase activity were evaluated . PLSR analysis revealed significant positive correlations ( = 0.5795) between the contents of triterpenes , , , and and the inhibitory effects of Alismatis Rhizoma. The present study establishes an effective method for simultaneous determination of multiple components, and identifies key bioactive triterpenes. These results can be used for systematic and novel analytical strategies for the quality control of Alismatis Rhizoma production.
ABSTRACT
Human cytosolic sulfotransferase 2A1 (SULT2A1) is an important phase II metabolic enzyme. The detection of SULT2A1 is helpful for the functional characterization of SULT2A1 and diagnosis of its related diseases. However, due to the overlapping substrate specificity among members of the sulfotransferase family, it is difficult to develop a probe substrate for selective detection of SULT2A1. In the present study, through characterization of the sulfation of series of bufadienolides, arenobufagin (AB) was proved as a potential probe substrate for SULT2A1 with high sensitivity and specificity. Subsequently, the sulfation of AB was characterized by experimental and molecular docking studies. The sulfate-conjugated metabolite was identified as AB-3-sulfate. The sulfation of AB displayed a high selectivity for SULT2A1 which was confirmed by reaction phenotyping assays. The sulfation of AB by human liver cytosols and recombinant SULT2A1 both obeyed Michaelis-Menten kinetics, with similar kinetic parameters. Molecular docking was performed to understand the interaction between AB and SULT2A1, in which the lack of interaction with Met-137 and Tyr-238 of SULT2A1 made it possible to eliminate substrate inhibition of AB sulfation. Finally, the probe was successfully used to determine the activity of SULT2A1 and its isoenzymes in tissue preparations of human and laboratory animals.
ABSTRACT
OBJECTIVE:To provide reference for confirming the protective effect of the establishment of PIVAS on antineoplastic drugs(ADs)occupational exposure to nursing staff in clinical departments,and to provide the basis for the formulation of ADs dispensing guideline and occupational exposure protection regulations. METHODS:By questionnaire survey and deriving data of lab examination index,the occurrence of abnormal menstruation,bad birth outcome,alopecia,blood toxicity, liver and renal toxicity in nursing staff of clinical departments with different ADs contact frequencies(non-exposure group as group A,low-exposure group as group B,high-exposure group as group C)were investigated and analyzed in our hospital before and after the establishment of PIVAS. The residual of ADs [methotrexate(MTX)and 5-Fluorouracil(5-FU)] in PIVAS environment were investigated by HPLC. RESULTS:A total of 160 questionnaires were sent out before the establishment of PIVAS,and 151 were effectively collected with effective recovery of 94.38%. After the establishment of PIVAS,150 questionnaires were sent out,and 144 were effectively collected with effective recovery of 96.00%. Questionnaire results showed that the incidence of abnormal menstruation,abnormal menstruation period,dysmenorrhea,spontaneous abortion,infertility and offspring low birth weight,the severity of hair loss in group C were significantly higher than group A,with statistical significance(P<0.05 or P<0.01). The incidence of above 6 conditions in group C were 5.14,6.10,3.81,4.04,6.15,8.08 times higher than in group A.At same time, the incidence of the offspring low birth weight and the severity of hair loss in group B were significantly higher than group A,with statistical significance(P<0.05 or P<0.01);the incidence of the former in group B was 6.21 times higher than in group A.After the establishment of PIVAS,the incidence of abnormal menstruation,abnormal menstruation period,dysmenorrhea,spontaneous abortion,infertility and offspring low birth weight,the severity of hair loss were decreased significantly in group C,with statistical significance(P<0.05). The severity of hair loss in group C was significantly higher than group A,and there was no statistical significance in above indexes,compared with group A(P>0.05). At the same time,there was no statistical significance in the incidence of the offspring low birth weight and the severity of hair loss between group B and A(P>0.05). Results of lab examination index investigation showed that before the establishment of PIVAS,WBC and PLT of group C were significantly lower than group A,and the incidence of abnormal liver function was significantly higher than group A,with statistical significance(P<0.05 or P<0.01). After the establishment PIVAS,WBC,PLT and RBC of group C,and PLT of group B were increased significantly compared to before the establishment PIVAS;the incidence of abnormal liver function in group C was decreased significantly compared to before the establishment PIVAS,with statistical significance(P<0.05 or P<0.01). There was no statistical significance in above indexes between group C and A(P>0.05),and PLT of group B was even significantly higher than group A(P<0.05). Results of investigation of ADs residues in PIVAS environment showed that there were different degrees of MTX and 5-FU residue on the surface of different objects. The residues of ADs from high to low were biological safety cabinet worktops,the floor just below biological safety cabinet,transfer box,transfer window and door handle and infusion bags. CONCLUSIONS:Nursing staff of clinical department with high ADs contact frequency face higher relevant health risks. The establishment of PIVAS can effectively protect the ADs occupational exposure of nursing staff in clinical departments,thereby reducing the above risks.At the same time,there are still different degrees of ADs residues on the surface of different objects in the PIVAS environment,and transshipment out of PIVAS is possible. It is suggested that the awareness of protection against occupational exposure risk caused by ADs residues in PIVAS environment should be improved;unified guideline for ADs dispensing and occupational exposure protection regulations should be formulated as soon as possible,so as to reduce occupational exposure risk associated with ADs for nursing staff in PIVAS and clinical departments.
ABSTRACT
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.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the microbiological transformation of paeoniflorin and albiflorin.</p><p><b>METHOD</b>The bacteria strains able to transform paeoniflorin and albiflorin were screened from 18 strains of microorganisms. The products were isolated by chromatography method and their structures were elucidated by spectral technology.</p><p><b>RESULT</b>It was found that Cunninghamella blakesleana (AS 3.970) and Syncephalastrum racemosum (AS 3.264) could convert paeoniflorin and albiflorin efficiently, respectively. C. blakesleana could convert paeoniflorin to produce albiflorin, while S. racemosum could convert albiflorin to produce paeoniflorin.</p><p><b>CONCLUSION</b>Paeoniflorin and albiflorin could be converted each other in definited condition.</p>