Objective: To study the in vitro antilipid-peroxidation effects of tannins extract from Sanguisorba officinalis on heart, liver, brain and kidney of rats.Methods: The in vitro inhibitory effects of tannins extract from Sanguisorba officinalis on lipid-peroxidation of heart, liver, brain and kidney of rats induced by Fe2+-cysteine (Fe2+-Cys), Fe2+-vitamin C (Fe2+-Vit C) and Fe2+-H2O2 were determined by spectrophotometry.Results: The inhibitory effects of tannins extract from Sanguisorba officinalis on MDA produced by lipid-peroxidation of brain, heart, liver homogenate, kidney and mitochondria of rats induced by Fe2+-Cys, Fe2+-Vit C and Fe2+-H2O2 were all significant.Conclusion: Tannins from Sanguisorba officinalis has good in vitro protective effects on antilipid-peroxidation of heart, liver, brain and kidney of rats, which is worth studying further.

Objective To investigate autophagy and proteasome system alteration in vivo and in vitro of diabetic nephropathy (DN) model rats.Methods Rat glomerular mesangial cells were primaryly cultured,and cell proliferation was tested by MTT assay.The mesangial cells were cultured under different concentrations of glucose (5.4 mmol/L for normal control and 30 mmol/L for high glucose) for 0,8,16,72 hours.The expression of autophagy (LC3) and proteasome (PSMAs) proteins was examined by Western blotting analysis.Spontaneous type 2 diabetes model OLETF and its normal control LETO rats were observed for 36 weeks.The levels of blood glucose and 24 hours urinary protein were evaluated in every 4 weeks.All the rats were sacrificed at the 36th week,and renal pathological changes were semi-quantitively analyzed.The expression of PSMAs and LC3 proteins was also examined in kidney cortex by Western blotting.Results Under high glucose concentrations,the abundance of PSMAs and LC3 proteins significantly reducedin the mesangial cells at 8 hours.There was no significant difference at other time points.The levels of blood glucose and 24 h urinary protein in OLETT rats exhibited progressive increase compared to those in LETO rats (all P＜0.01).And glomerular sclerosis index and tubulointerstitial injury index were significantly higher than those in LETO rats (all P＜0.01).The abundance of PSMAs proteins was significantly reduced in renal cortex of OLETF rats compared with LETO rats,while the abundance of LC3 proteins had no significant difference between two groups.Conclusion Proteolytic system dysfunction may play a role in pathogenesis of DN.

Objective:To study the effect of tannins from Pericarpium Granati on hepatic microsomal enzyme in rats. Methods:Thirty Wistar rats were randomly divided into five groups:the blank control group, high, medium and low dose groups of tannins from Pericarpium Granati, the positive control group of phenobarbital sodium. The blank control group was given physiological saline. The three different doses groups were respectively given tannins from Pericarpium Granati orally at the dose of 150,100 and 75 mg·kg-1 · d-1 for 7 days. The positive control group was given phenobarbital sodium 80 mg·kg-1 ·d-1 with intramuscular injection for 5 days. At the end of the experiment, the activity ofⅠandⅡphase metabolic enzymes in liver microsomes of each group was determined by UV. Results:Compared with the blank control group, the high, medium and low dose groups of tannins from Pericarpium Granati could sig-nificantly decrease the content of CYP 450 and CYPb5, and inhabit the activity of ADM (P<0. 01);the high and medium dose group could significantly inhibit ERD enzyme activity (P<0. 01);the high dose group could significantly reduce GST enzyme activity (P<0. 01). Conclusion:Tannins from Pericarpium Granati has notably inhibitory effect on hepatic microsomal enzyme in rats, which can reduce the expression of CYP3A and CYP2E1 in a dose-dependent manner.

Objective The feasibility of predicting the B-cell epitopes of human Neutrophil Gelatinase-Associated Lipocalin (NGAL) was discussed by applicating bioinformatics technology.Linear epitope molecules that have diagnostic value were screened and these recombinant linear multi-epitope peptides were constructed,and expressed.The immunogenicity of the recombinant linear multi-epitope peptides were also identified.Methods NGAL amino acid sequence was got from GenBank in the Department of Clinical Laboratory of the Second Affiliated Hospital of Nanjing Medical University in July 2015,the Predicted,ABCpred,BepiPred,BcePred,and Lasergene softwares were used to predict the linear B cell epitope prediction.The predict epitopes were constructed and prokaryotic expressed,and then the single epitope antigens which could reacted with commercially available polyclonal NGAL antibody were screened out by Western blot.Finally,the multi-epitope peptide was constructed,expressed,and identified through immunoreactions.Results Eight possible epitopes were obtained after prediction.pET32a-N1-N8 prokaryotic expression vector were used to express the predict epitopes.After purification and Western blot analysis,three of the epitopes have strong antigenicity,and then a soluble fusion protein was expressed and obtained from the multi-epitope prokaryotic expression vector pET22b-Ngal_MEP1.The fusion protein was successfully purified by Ni2 + affinity column.Western blot analysis showed that the fusion protein had a strong antigenicity.Conclusions The constructed multi-epitope linear NGAL antigen peptides can obtain high soluble expression in prokaryotic expression system,and have a strong immunoreactivity,which can be used in subsequent antibody preparation.

Objective:To investigate the transport mechanism of punicalagin in MDCK monolayer model .Methods:The safe con-centration of punicalagin in MDCK cells was determined by CCK8 assay.Millicell -ERS was used to measure cell monolayer TEER value to determine the integrity of the cell monolayer .The effects of direction , drug concentration , time, P-gp inhibitor and EDTA-Na2 on the absorption and transport of punicalagin were studied systematically .And then the drug concentration was analyzed by HPLC to calculate the apparent permeability coefficient (Papp) and efflux ratio(ER).Results: Punicalagin transport in MDCK cells was time and concentration dependent .Punicalagin showed poor absorption in MDCK cells .Papp from apical to basolateral side ( AP-BL) within the concentration range of 100-300μg· ml-1 was (6.13 ±0.12) ×10 -7 cm· s-1 , (6.96 ±0.26) ×10 -7 cm· s-1 and (5.94 ±0.10) ×10 -7 cm· s-1 , respectively .P-gp inhibitor and EDTA-Na2 could significantly increase the transport of punicalagin in AP-BL direc-tion, while the transport decreased at 4℃.Conclusion:The transport mechanism of punicalagin might be passive diffusion as the dom-inating process involving active transportation .Punicalagin is one of P-gp substrates with exocytosis and absorbed via the paracellular route.

ABSTRACT Ellagitannin is a complex compound of plant polyphenols, and it is an effective substance in many commonly used natu-ral drugs. Ellagitannin has been widely used in medicine, cosmetics, food, health care products and the other fields. The article re-viewed the structure source, biological activity and metabolic outcomes of ellagitannin in order to provide basis for the in-depth resear-ches and the resource development of natural medicines rich in tannins.

ABSTRACT Ellagitannin is a complex compound of plant polyphenols, and it is an effective substance in many commonly used natu-ral drugs. Ellagitannin has been widely used in medicine, cosmetics, food, health care products and the other fields. The article re-viewed the structure source, biological activity and metabolic outcomes of ellagitannin in order to provide basis for the in-depth resear-ches and the resource development of natural medicines rich in tannins.