Objective To explore the effect of sodium arsenite (iAs3+) and sodium hydrogen arseuate (iAs5+) exposure on rat multidrug resistance protein 2 (MRP2) expression,and the correlation of liver arsenic and its methylation products,and to provide a basis for further elucidating the mechanism of arsenic toxicity.Methods Seventy wistar rats were randomly divided into seven groups,10 rats in each group.Control group was administrated with deionized water.Sodiun arsenite high-dose group,middle-dose group,low-dose group were administrated with different concentrations of sodium arsenite:20.0,6.7 and 2.2 mg/kg BW every day,respectively.Animals were sacrificed 90 days later by cervical dislocation to collect liver,the expression of MRP2 in the membrane of hepatocyte was determined by Western blotting.HPLC-HGAFS was employed to determine the content or level of arsenic and its methylation products.Correlation between expression of MRP2 and arsenic methylation products was analyzed using Pearson's linear correlation method.Results The MRP2 protein levels of control group,iAs3+ and iAs5+ high-,middle-,low-dose groups were 1.21 ± 0.13,1.85 + 0.09,1.65 + 0.19,1.61 + 0.18,1.69 + 0.04,1.42 + 0.19,1.27 ± 0.10.Compared to control group,MRP2 protein levels of iAs3+ high-,middle-,low-dose group and iAs5+ high-dose group were increased (P ＜ 0.05); Compared with the low-dose group,MRP2 protein levels of iAs3+ and iAs5+ high dose groups were increased(P ＜ 0.05); Comparing the matched doses group of iAs5+ and iAs3+,MRP2 protein levels of iAs3+ high-,middle-,low-dose group were higher than iAs5+(P ＜ 0.05).Meanwhile,there was a significant positive relationship between the expression of MRP2 and the content of iAs3+,MMA and DMA in iAs3+ exposed group(r =0.575,0.678,0.395,all P ＜ 0.05).There was also a significant correlation between the expression of MRP2 and MMA and IMA in iAs5+ exposed group(r =0.593,0.643,all P ＜ 0.05).There was no significant relationship between the expression of MRP2 and the content of iAs5+in iAs5+ exposed group.Conclusions The expression of MRP2 is increased with increasing dose of arsenic exposure,thus resulting in compensatory changes on MRP2 expression in the liver cell membrane.MRP2 efflux may play an important role in the metabolic pathways of iAs3+.The level or load of arsenic methylation in liver is closely related with MRP2 expression.

1H NMR-based metabolomic approach combined with multivariate statistical analysis was used to evaluate the quality of 21 Farfarae Flos (FF) samples from different growth regions. Principal component analysis showed that wild and cultivated FF could be separated clearly, suggesting a big chemical difference existed between them. Supervised PLS-DA analysis indicated that the wild samples showed higher levels of secondary metabolites, such as bauer-7-ene-3β, 16α-diol, chlorogenic acid, rutin, 7-(3'-ethylcrotonoyloxy)-1α-(2'-methyl-butyryloxy)-3, 14-dehydro-Z-notonipetranone (EMDNT), tussilagone, β-sitosterol and sitosterone. This is consistent with traditional experience that the quality of wild samples are better than that of cultivated ones. The content of pyrrolizidine alkaloids senkirkine also differed greatly among samples from different habitats. The Pearson correlation analysis showed that senkirkine is positively correlated with 4, 5-O-dicaffeoylquinic acid, 3,5-O-dicaffeoylquinic acid, 3,4-O-dicaffeoylquinic acid, rutin, kampferol analogues, to a statistically significant extent. The correlation between the toxic compounds and the bioactive components in FF should be further studied.
Chlorogenic Acid ; Drugs, Chinese Herbal ; chemistry ; Flowers ; chemistry ; Magnetic Resonance Imaging ; Magnetic Resonance Spectroscopy ; Metabolomics ; Quinic Acid ; analogs & derivatives ; Rutin ; Sitosterols ; Tussilago ; chemistry

Objective Helicobacter pylori（ ）， To investigate the infection status of HP and analyze the correlation between HP Methods infection and serum bilirubin in railway drivers. A total of 2 731 railway drivers in Zhengzhou locomotive depot were - selected as study subjects using judgment sampling method. Carbon 13 urea breath test was used to evaluate the HP infection ， status. The metabolic indexes of HP positive group and HP negative group were compared and the relationship between HP Results （ ） ， infection and serum bilirubin was analyzed. The HP infection rate was 42.3% 1 156/2 731 . The older the age the ， （ ）， （ P ） longer the work years and the higher the body mass index BMI the higher the HP infection rate all <0.01 . The infection （P ） rate of HP in married people was higher than that in unmarried people <0.01 . The HP infection rate of smokers was higher - （P ） - ， than that of non smokers <0.01 . Compared with the HP negative group fasting blood glucose and serum levels of total ， （ - ）， （ ） - cholesterol low density lipoprotein cholesterol LDL C triglyceride and homocysteine Hcy were increased in the HP （ P ） （ - ）， ， positive group all <0.05 . The serum levels of high density lipoprotein cholesterol HDL C total bilirubin direct bilirubin （ ） - （ P ） DBIL and indirect bilirubin were lower than those in HP negative group all <0.05 . Logistic regression analysis showed that （ P ） HP infection was associated with low serum total bilirubin and low DBIL all <0.01 after adjusting for the confounding effects ， ， ， ， ， ， ， - ， - ， of age work years marital status smoking history fasting blood glucose total cholesterol triacylglycerol LDL C HDL C Conclusion ， ， ， and Hcy. The age work length BMI smoking and marital status are the influencing factors of HP infection in railway drivers. HP infection is associated with low levels of total bilirubin and DBIL.

Four salts of ticagrelor, ticagrelor-3,5-dinitrobenzoic acid, ticagrelor-pyrazinamide, ticagrelor-D-proline and ticagrelor-L-proline were prepared by solvent suspension and liquid-assisted grinding to improve the solubility of ticagrelor. The compounds were characterized by powder X-ray diffraction, Fourier transform infrared spectroscopy, differential scanning calorimetry, nuclear magnetic resonance spectroscopy, elemental analysis, and the intermolecular salt-bonding forces were analyzed. The equilibrium solubility of salts and pure drug in hydrochloride buffer pH 1.2 and phosphate buffer pH 6.8 were measured by high-performance liquid chromatography. Ticagrelor was salted with 3,5-dinitrobenzoic acid, pyrazinamide, D-proline, L-proline all in a stoichiometric ratio of 1∶1; with the exception of ticagrelor-D-proline, the solubility of the other three salts provided significantly improved solubility in hydrochloride buffer pH 1.2, and the equilibrium solubility of ticagrelor-3,5-dinitrobenzoic acid was increased by approximately 1.7 folds as compared to pure drug. Salt-forming technology is convenient and can improve the solubility of ticagrelor.

Objective: To study the effect of and possible mechanism of knockinng down PI3Kp85α using siRNA in MCF-7 human breast cancer cell line. Methods: Oligofectamine was used to transfect PI3Kp85α siRNA to knock down the PI3Kp85α expression level in MCF-7 human breast cancer cell line in vitro. Real-time PCR was conducted to detect the expression of PI3Kp85α. The effect of PI3Kp85αsiRNA on the growth of MCF-7 cells was measured by MTT. The cell cycle distribution and cell apoptosis were detected by cell flow cytometry. Protein expression was evaluated by immunofluorescence staining and Western blot. Results: The expression of PI3Kp85 α was knocked down with PI3Kp85α siRNA in MCF-7 cells. Cell growth was delayed in PI3Kp85αsiRNA-treated group. Conclusion: The suppressive effect of PI3Kp85αsiRNA on the growth of MCF-7 human breast cancer cell line is significant and PI3Kp85α could be a candidate for gene therapy for breast cancer.

A multiplex real-time PCR was developed to detect ctxA of Vibrio cholerae, gyrB and tdh of Vibrio parahaemolyticus simultaneously. The multiplex real-time PCR were evalidated by detection for the three genes in 47 toxigenic V. cholerae O1 and O139 strains (ctxA+; O1=3, O139=44), 25 non-toxigenic V. cholerae strains (ctxA-; O1=12, O139=6, non-O1 and non-O139=7), 116 V. parahaemolyticus strains with or without tdh (73 or 43) and 9 other bacteria strains. The specificity and sensitivity of the multiplex real-time PCR in detection for the ctxA and the tdh genes in the strains tested were both 100.0%, compared to the results by routine PCRs. In the detection for V. parahaemolyticus specific gyrB using the multiplex real-time PCR, all of 116 V. parahaemolyticus strains were positive, and 9 other strains and 72 V. cholerae strains were all negative. The multiplex real-time PCR is a sensitive, specific and quick assay not only for detecting virulence genes of V. cholerae and V. parahaemolyticus but also for identifying V. parahaemolyticus at species level. In addition, two real-time PCRs for detection of V. parahaemolyticus virulence genes trh1 and trh2 were also developed.

Objective To determine the value of Magnetic resonance imaging(MRI)technique on diagnosis of adrenal medulla hyperplasia model rats confirmed by pathology study.Methods Sixty male SD rats were divided into 6 groups randomly,10 rats in each group.As experiment group,A,B,C groups were subcutaneously injected with reserpine(0.4 mg?kg~(-1)?d~(-1))from the beginning to 40,60,80 days respectively.As compare,the control groups(a,b,c group)were only injected with sodium chloride simultaneously.MRI technique and pathology study were performed for all the subjects on 40,60,and 80 days respectively.Results The percent of medulla were higher in B and C groups than that in b and c groups respectively[(34.3?5.8)% vs.(25.7?8.9)%,t=2.462,P

Objective To investigate the effects of neutrophil gelatinase associated lipocalin(NGAL) on renal tubular epithelial cells apoptosis and apoptosis-regulated protein fas,bcl-2 in rat renal ischemia reperfusion injury (IRI).Methods Renal IRI models of rats were established.Rats were randomly divided into 3 groups,including control group,IRI model group and NGAL group.The pathological change of kidney tissue was investigated by hemotoxylin-eosin staining.Renal tubular epithelial cells apoptosis was detected by TUNEL method.Expression of fas and bcl-2 was measured by real-time PCR and Western blotting.Results Compared with IRI model group,NGAL group showed a decreased number of renal tubular epithelial cells apoptosis [(8.6±3.4)/HP vs (20.8±3.7)/HP,P＜0.05],down-regulated fas mRNA (2.34±0.51 vs 6.84±2.34,P＜ 0.05),fas protein (0.65±0.05 vs 0.95±0.08,P＜0.05) and up-regulated bcl-2 protein (0.33±0.05 vs 0.24±0.03,P＜0.05),but the bcl-2 mRNA had no significant change.Conclusion NGAL can protect renal tubular epithelial cells in renal IRI,which may be associated with decreasing cell apoptosis and adjusting protein expression by apoptosis-regulated cytokines.

Objective To explore the molecular mechanism of renal function defects after urinary obstruction and investigate the effect of sirolimus on the expression of γ-ENaC,Na + K + ATPase and AQP2,and its mechanism of renal Water-Electrolyte imbalance following bilateral ureteral obstruction (BUO) in rat kidneys.Methods Forty-eight rats were randomly divided into a sham operation group ( sham group),a BUO group,and a sirolimus treatment after BUO group.Bilateral ureters were exposed and occluded with ligature in the BUO and sirolimus treatment groups.Twenty-four hours later,the obstructed ureters were decompressed by removal of the ligature.The sham animal group underwent identical surgical procedures,but the ureter was simply dissected without removal of the ligature.The sirolimus treatment groups was given sirolimus intragastricly 0.4 ml per day (2 mg/kg · d) from the day before surgery until the rats were scari fled.The sham and BUO groups were given the same volume of intragastric saline.The urine and blood were collected at 4 d,7 d after surgery,and the functional data were observed.The expression of γ-ENaC,Na+K + ATPase and AQP2 were examined by immnohistochemistry and immunoblotting.Results On day four and seven post ureteral obstruction release,urine volume in the BUO group were (85.31 ± 13.15,66.39 ±10.56 ml),significantly higher than that of sham operation (35.36 ± 7.74,33.90 ± 8.03 ml) and sirolimus treatment groups (69.81 ± 10.70 ml,48.57 ± 9.01 ml) (P ＜ 0.05 ).Urine sodium concentrations in the BUO group were (42.17 ± 7.35 mmol/L,43.63 ± 18.39 mmoL/L),significantly lower than that of sham operation ( 170.56 ± 18.39 mmoL/L,172.52 ± 7.35 mmol/L) and sirolimus treatment groups (76.18 ± 13.20 mmol/L,134.28 ± 13.20 mmol/L),P ＜ 0.05.Immunoblotting assay showed that,on day four and seven post rats ureteral obstructions were released,integral optical density of γ-ENaC (2.09 ±0.32,2.27 ±0.35),Na+ K+ATP enzyme (2.41 ±0.48,2.67 ±0.43) and AQP2 (2.17 ±0.45,2.63 ±0.28) in the sirolimus treatment group were significantly higher than those of BUO group ( 1.28 ± 0.21,1.45 ±0.17) (1.99 ±0.28,2.18±0.24) (0.93 ±0.22,1.31 ±0.16),but still lower than the sham group (2.58±0.51,2.60±0.56) (2.89±0.53,2.97 ±0.66) (3.05 ±0.63,3.10±0.67).There were significant differences among all the three groups ( P ＜ 0.05 ).Conclusions The downregulation of γ-ENaC,Na + K + ATPase and AQP2 expression after BUO may contribute to the impaired renal tubular sodium reabsorption,decreased urinary concentration,and postobstructive polyuria.Sirolimus treatment significantly prevents impairment in renal function and also prevents downregulation of y-ENaC,Na + K+ ATPase and AQP2during BUO,demonstrating a marked renoprotective effect of sirolimus treatment in conditions with urinary tract obstruction.

AIM To establish an HPLC-ELSD method for the simultaneous content determination of gastrodin,parishin A,parishin B,ginsenosides Rg1,Re,Rf,Rb1 and Rd in Tiantai No.1 Tablets (Ginseng Radix et Rhizoma,Gastrodiae Rhizoma,Cistanches Herba,etc.).METHODS The analysis of methanol extract of this drug was performed on a 45 ℃C thermostatic Alltima C1scolumn (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrie-0.1％ phosphoric acid flowing at 1.0 mL/min in a gradient elution manner.RESULTS Eight constituents showed good linear relationships within their own ranges (r ＞ 0.999 0),whose average recoveries were 96.94％-97.93％ with the RSDs of 1.06％-2.48％.CONCLUSION This simple,accurate and reliable method can be used for the quality control of Tiantai No.1 Tablets.