The study aims to establish a quantitative nuclear magnetic resonance (QNMR) method for the determination of the absolute content of bosentan. Proton nuclear magnetic resonance spectroscopy [1H NMR] spectra were obtained in CDCl3 with the internal standard dimethyl terephthalate and zg30 pulse sequence by using a Bruker AVANCE II 400 spectrometer. The content of bosentan is determined with QNMR in comparison with the result obtained by mass balance method. The result is 96.25% by QNMR and 96.54% by mass balance method. A rapid and accurate QNMR method has been established for the quantitative determination of the absolute content of bosentan. The study provides a new way for the quality control and calibration of a new reference standard material, it could be the complementary with the mass balance method for the assay of standard reference.
Calibration ; Magnetic Resonance Spectroscopy ; methods ; Molecular Structure ; Protons ; Quality Control ; Sulfonamides ; chemistry

Objective To investigate the correlation between chronic hepatic diseases and small intestinal inflammation.Methods Patients who received capsule endoscopy in The First Affiliated Hospital of Guangdong Pharmaceutical University were divided into groups of liver cirrhosis,non-alcoholic fatty liver disease(NAFLD),chronic hepatitis and non-hepatic disease according to clinic data from August 2011 to August 2015.The severity of small intestinal mucosal inflammation was graded according to Lewis Scoring system and incidence of small intestinal lesions in different groups and Lewis scores were compared.The liver function was also graded with liver noninvasive scoring systems.Then the correlation between liver function damage and small intestinal lesions was investigated.Results A total of 338 cases were enrolled in the study,including 25 cases of liver cirrhosis,47 cases of NAFLD,20 cases of chronic hepaitis and 246 cases of non-hepatic disease.There were 22 (88.0％),36 (76.6％),12 (60.0％) and 78 (31.7％) cases with lesions in small intestine in the four group respectively with significant differences(P＜0.001).Rate of small intestinal villi edema was significantly higher in liver cirrhosis group,NAFLD group,chronic hepatitis group than that in non-hepatic disease group(all P＜0.017).Small intestinal villi edema was found mainly in the upper and one third of middle parts in small intestine (P =0.033).Lewis scores of liver cirrhosis group (190.80±228.42)and NAFLD group(125.38± 191.31) were higher than those of non-hepatic disease group (42.91±97.69,P=0.021,P =0.034).Forns score,FIB-4 score,NAFLD-FS score and Child-Pugh score were positively correlated with Lewis score (correlation coefficient:0.247,0.244,0.223,0.284respectively,all P＜0.001).Conclusion Chronic hepatic diseases such as liver cirrhosis,NAFLD,chronic hepatitis might be risk factors for small intestinal mucosal inflammation,and the severity of chronic hepatic diseases may be positively correlated with that of small intestinal mucosal lesions.

Objective To evaluate the clinical value of liquid-based thinprep cytology test (TCT) and human papillomavirus (HPV) in cervical lesions diagnosis.Methods 525 patients with TCT was inflammation and abnormalities had HPV testing and colposcopy with biopsy analysis.Results Histopathological diagnosis as CIN Ⅰ or CIN Ⅰ above cases percentage of TCT of ASCUS,LSIL,HSIL,and cancer cases were 55 ％ (160/290),88 ％ (99/113),92 ％ (24/26) and 100 ％ (4/4),Pathological diagnosis as inflammation and glandular phosphate in 441 cases of this TCT-positive patients were 148 patients,false positive 34 ％.TCT-negative patients,pathologically confirmed of CIN in 84 cases patients with moderate to severe cervical erosion or cervical polyps were 22 cases,26 ％ TCT false negative.They had statistically significance (x2 =815.4,P =0.000).74 ％ HPV positive patients diagnosed as CIN Ⅰ or CIN Ⅰ above (x2 =104.94,P =0.000).Diagnosed as CIN Ⅰ cases in less than 30 years old accounted for 62 ％ (61/98),CIN Ⅱ and above for only 38 ％ (37/98) and without cancer.But the CIN Ⅰ in above 30 age group was significantly reduced to only 37 ％ (81/217),CIN Ⅱ and above was significantly increased to 63 ％ (136/217),and cancer rate increased (x2 =63.71,P =0.012).Histopathological diagnosed as CIN Ⅰ case were 26 ％ (83/315) of cervical smooth,74 ％ (232/315) of cervical erosion; 14 cases of cancer occurred in patients with cervical erosion.It was statistically significant (x2 =39.9,P =0.014).Conclusion TCT and HPV for screening of cervical lesions play an important role in cervical lesions screening.

Objective To observe the change of intestinal microflora on the process of nonalcoholic steatohepatitis(NASH),and to explore the synbiotics therapeutic effect on NASH.Methods Rats were administrated with high fat diet to establish NASH model.In the process of NASH rats modeling,the level of triglyceride (TG), total cholesterol (TC), high-density lipoprotein (HDL), low density lipoprotein (LDL), fasting blood sugar (FBS) and fasting insulin (FINS) was dynamically tested by automatic biochemical analyzer.The change of main intestinal flora was detected by 16 S rRNA fluorescence quantitative polymerase chain reaction.NAFLD activity score was calculated.HE staining was used to observe the hepaticpathological changes and the TLR4 expression was detected by using enzyme-linked immunosorbent assay and immunohistochemical method.Until the 4th,8th,10th weekin the process of NASH modeling, 10 rats were feeded with synbiotics for 2 weeks, and all of above indicators were tested and observed.Results 1)With the extension of a high-fat diet feeding time, the degree of hepatocyte steatosis obviously increased.NAFLD score was significantly heightened(P<0.01).2)Number of independent activities of rats significantly increased, the serological level of TG, TC, LDL, FBS and FINS were lower significantly after intervention with synbiotics for 2 weeks(P<0.05).3)Synbiotics intervention for two weeks significantly increased the amount of bifidobacterium and lactobacillus and decrease the amount of enterococcus significantly(P<0.05).4)The expression of TLR4 was gradually increased in the process of NASH rats modeling(P<0.05),but decreased after 2 weeks of the synbiotics-intervention (P<0.05).Conclusions Intestinal microecology change is closely related to the development of NASH,therefor, synbiotics could improve the quality of life and biochemical indicators of NASH rats through adjusting intestinal microecology and the expression level of TLR4 protein might been involved.

Objective To in ve stigate the effects of advanced glycation end products (AGEs) on matrix metallop roteinase-2 (MMP-2) activity and its mRNA expression in renal cortex of rats. Methods Diabetic model of rats was induced by s treptozotocin. AGEs were prepared by incubation of rat serum protein with 0.5 mo l/L glucose. AGEs was administered intravenously to normal rats (AGEs group), an d native rat serum protein was given as negative control (negative group) and no rmal rats without treatment were as control (control group). AGEs content in ren al cortex and serum was quantified by ELISA, MMP-2 and TIMP-2 mRNA expressions were examined by RT-PCR and MMP-2 activity was measured by zymography. Results AGEs content increased significantly, MMP-2 mRNA expression descended and TIMP-2 mRNA expression ascended in renal cortex o f diabetic rats (all P

OBJECTIVE:To provide reference for improving the quality of talent training for medical direction of chain man-agement major in higher vocational college. METHODS:The situation of pharmaceutical retail chain industry market was analyzed in China;the problems of talent training mode were summarized to put forward reform plan and measures according to disadvantag-es. RESULTS:Based on the market situation as continues expansion of pharmaceutical retail chain industry market scale,shortage of professional talent pool,the formulation of new retail mode in China,and training mode situation as not enough in-depth cooper-ation between college and enterprise,poor practicality and pertinence of course setting,irrational teachers'structure,teaching re-form could be conducted on the basis of modern apprenticeship system in following aspects,such as college and enterprise shared the responsibility of training and cultivating,student admission is recruitment;college and enterprise designed curriculum system and assessed students together;double tutorteam of college teacher and enterprise teacher was constructed. CONCLUSIONS:The talent training mode based on modern apprenticeship system is the entry point for teaching reform of medical direction of chain management major in higher vocational college,which is conducive to cultivate high quality pharmaceutical management talents meeting industry needs and social development.

AIM: To explore the influence of CD47 molecules on the maturation and function of cultured dendritic cells (DCs). METHODS: Monocyte cell-derived DCs were propagated in granulocyte-macrophage colony-stimulating factor (GM-CSF) and lipopolysaccharide (LPS) plus interleukin (IL)-4, in the presence or absence of anti CD47 monoclonal antibodies (anti-CD47 mAbs). Flow cytometry was used to detect the cell surface phenotype. The concentration of IL-12P70 in supernatant was measured by ELISA technique. The antigen-presenting functions of DCs were determined in one-way mixed leukocyte reaction by Brdu-ELISA. Electrophoretic mobility shift assays (EMSA) was used to examine NF-?B activity. RESULTS: The anti-CD47 mAbs markedly suppressed the expression of CD80,CD86,CD83,CD1a,HLA-DR on the surface of DCs (P

Objective To investigate the possible contributing risk factors, diagnosis and treatment of gastroparesis after abdominal surgery. Methods Potential risk factors, clinical manifestations and therapeutic approaches of 22 cases of gastroparesis after abdominal operation were analyzed retrospectively. Results In this series, gastroparesis more frequently occured in those patients having a long-term preoperative gastric outlet obstruction, malignant tumor, O type blood group pancreaticoduodenectomy or simple gastrojejunostomy without gastric resection. Endoscopy, upper gastrointestinal radiography and radioisotope scintigraphy were effective methods in the diagnosis of gastroparesis, and normal gastrointestinal movement was achieved mostly in 5 weeks postoperatively by conservative treatments.Conclusions Postoperative gastroparesis can recover spontaneously after a longer period of supportive treatment combined with prokinetic drugs, and reoperation should be avoided.

AIMTo elevate the encapsulation efficiency, decrease the burst release and improve the release of protein entrapped in poly (lactic-co-glycolic acid) (PLGA) microspheres. The bovine serum albumin (BSA) composite microspheres of alginate-chitosan-PLGA were prepared and the release characteristics of BSA from this composite microspheres were studied.

METHODSThe much smaller calcium alginate microcapsules were first prepared by a modified emulsification method in an isopropyl alcohol-washed way and coated with chitosan, then the alginate-chitosan microcapsules were further entrapped in PLGA to form the composite microspheres. The protein concentration was determined using a BCA protein assay kit. The release profiles were changed with various formulation factors.

RESULTSThe average diameter of the composite microcapsules was about 30 microm. Comparing with 60% to 70% of the conventional PLGA microspheres, the average encapsulation efficiency was more than 80%, and the burst releases in phosphate buffer solution of the composite microspheres decreased from 40% and 50% to 25% and further to 5% in saline solution.

CONCLUSIONThe novel composite microspheres were prepared, the drug encapsulation efficiency increased and the burst release decreased. The desired release profiles could be obtained by regulating the ratios of PLG and PLA in the composite microspheres.

Alginates ; chemistry ; Chitosan ; chemistry ; Drug Delivery Systems ; methods ; Lactic Acid ; chemistry ; Microspheres ; Particle Size ; Polyglycolic Acid ; chemistry ; Polymers ; chemistry ; Serum Albumin, Bovine ; chemistry

Sphingosine kinase 1 (SphK1) plays critical roles in cell biological functions. Here we investigated the effects of SphK1 inhibitor SKI II on hepatoma HepG2 cell cycle progression and invasion. Cell survival was determined by SRB assay, cell cycle progression was assayed by flow cytometry, the ability of cell invasion was measured by Matrigel-Transwell assay and protein expression was detected by Western blotting. The results showed that SKI II markedly inhibited HepG2 cell survival in a dose-dependent manner, induced G1 phase arrest in HepG2 cell and inhibited cell invasion. SKI II markedly decreased the expressions of G1-phase-related proteins CDK2, CDK4 and Cdc2 and the levels of cell invasion-associated proteins MMP2 and MMP9. The results showed that SKI II inhibited cell cycle progression and cell invasion, implying SphK1 as a potential target for hepatoma treatment.
CDC2 Protein Kinase ; Cell Movement ; drug effects ; Cell Survival ; drug effects ; Cyclin-Dependent Kinase 2 ; metabolism ; Cyclin-Dependent Kinase 4 ; metabolism ; Cyclin-Dependent Kinases ; metabolism ; G1 Phase ; drug effects ; Hep G2 Cells ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Phosphotransferases (Alcohol Group Acceptor) ; antagonists & inhibitors ; Thiazoles ; pharmacology