Objective To establish a RP-HPLC method for determining the concentration of prulifloxacin active metabolite in human plasma and urine.Methods The supernatant obtained by centrifugation after the sample was precipitated with methanol- acetonitrile (1:1) was chromatographically separated on a Diamonsil C_(18)(250 mm?4.6 mm,5?m) using a mobile phase con- sisting of acetonitrile and 0.05 mol/L potassium dihydrogen phosphate (pH2.2) containing 1% tetrabutylammonium bromide. The solutions of 20:80 (V/V) and 12:88 (V/V) at a flow rate of 1.0 mL/min and 1.6 mL/min were used for plasma and u- rine, respectively.Then the samples were assayed at wavelength of Ex 280 nm and Em 425 nm.Results The linear range for prulifloxacin active metabolite in plasma and urine were 0.005-5 mg/L (r=0.9999) and 0.05-5 mg/L(r=0.9999)with a low- er limit of quantitation of 0.002 mg/L and 0.01 rag/L, respectively.In plasma, the relative recovery ranged from 100.64% to 101.00% at the concentration of 5.00, 0.50 and 0.05 mg/L and within-day and between-day precisions were less than 2.5% and 4.6% respectively.Meanwhile, the relative recovery ranged from 97.20% to 100.20% at the concentration of 2.50, 0.50 and 0.10 mg/L in urine.The within-day and between-day precisions were lower than 1.3% and 4.3%, respectively.The method had been successfully used for the pharmacokinetic studies of a prulifloxacin formulation after oral administration to healthy volunteers.Conclusions The present method is simple, rapid, accurate, reproducible and suitable for the pharmacoki- netic study of prulifloxacin in humans.

Objective To evaluate the prognostic factors in locally advanced non-small-cell lung cancer (LA-NSCLC) for selectively carrying out prophylactic cranial irradiation (PCI).Methods 114 patients with LA-NSCLC between Jun 2006 and Oct 2010 were retrospectively analyzed. Related risk factors and features about brain metastases were analyzed.Results The 2-year incidence rate of brain metastases was 31.58 ％ (36/114),the first brain metastases was 20.18 ％ (23/114),and sole brain metastases was 9.65 ％(11/114),respectively.Variables involved in the equation of binary logistic regression analysis were pathology (OR =5.892) and treatment mode(OR =2.888).The incidence rate of brain metastases in patients of non-squamous carcinoma and single treatment mode was higher than others (P ＜ 0.01) Model fitting is better (P ＞ 0.05).Overall accuracy rate of predicting brain metastases is 67.7 ％.The increased rate of lactate dehydrogenase in the patients with brain metastases or death was 17.54 ％, which was higher than that in the survival patients without brain metastases (P ＜ 0.01).At the same time,the station number and the number of mediastinal lymph node metastases were positively correlated (r =0.716, P ＜ 0.01).The incidence rate of brain metastases or mortality rate was higher in the adenocarcinoma cases than that in the squamous carcinoma cases (P ＜ 0.01,P ＜ 0.05),with more frequent occurrence of mediastinal metastases.The mean diameter of squamous carcinoma and adenocarcinoma were 5.8 cm and 3.9 cm, respectively (P ＜ 0.01).Conclusions The incidence rate of brain metastases was higher in patients with single treatment.Large primary tumors, high lactate dehydrogenase, non-squamous carcinoma, multiple stations, and multiple mediastinal lymph nodes metastases can be regarded as risk factors of brain metastases to perform PCI.

Based on the results of the morphologic studies on genus Dioscorea, the paper summarized the entire chemical constituent that isolated from this genus and analyzed it with the methods of chemotaxonomy. The rules of the chemical constituent and pharmacodynamic effects were analyzed. Seventeen species which belong to Sect. Stenophora Uline of Dioscorea contain steroidal sapogenin. Other species with different main components such as polysaccharide and tannin have have different effects. This chemotaxonomic view point will conduce to establish a phylogeny of the genus Dioscorea.
Animals ; China ; Dioscorea ; chemistry ; classification ; genetics ; growth & development ; Humans ; Phylogeny ; Plant Extracts ; chemistry ; pharmacology

To figure out the stability and intestinal bacteria metabolites of rats in vitro of astragaloside IV ( AST), this research was done to explore the stability of AST in the artificial gastric juice. artificial intestinal juice and rat liver homogenate and the metabolism in rat intestinal in vitro. HPLC was used to calculate the remaining rate of AST in biological samples by measuring the content of AST, while metabolites were determined by combining the methods of TLC, HPLC and LC-MS/MS. It turned out that AST was difficult to metabolize in the artificial gastric juice, artificial intestinal juice and rat liver. Also, the metabolic pathway of AST was stepped by deglycosylation. Firstly, AST was converted to its secondary etabolites (6-O-β-D-glucopyranosyl- cycloastragenol, CMG) by removal of xylose moiety at C-3, then transformed into cycloastragenol (CAG) after hydrolytic removal of the glucose moiety at C-6. All the results suggested that the metabolism of AST in vivo occurs mainly in the intestinal by hydrolysis of glycosyl. In conclusion, hydrolysis of intestinal flora is the main reason that AST metabolizes.
Animals ; Bacteria ; metabolism ; Chromatography, High Pressure Liquid ; Drug Stability ; Intestines ; microbiology ; Liver ; metabolism ; Rats ; Rats, Sprague-Dawley ; Saponins ; chemistry ; metabolism ; Tandem Mass Spectrometry ; Triterpenes ; chemistry ; metabolism

Epiberberine, one of the most important isoquinoline alkaloid in Coptidis Rhizoma, possesses extensive pharmacological activities. In this paper, the liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to study phase I and phase II metabolites. A Thermo HPLC system (including Surveyor AS, Surveyor LC Pump, Surveyor PDA. USA) was used. The cocktail probe drugs method was imposed to determine the content change of metoprolol, dapsone, phenacetin, chlorzoxazone and tolbutamide simultaneously for evaluating the activity of CYP2D6, CYP3A4, CYP1A2, CYP2E1 and CYP2C9 under different concentrations of epiberberine in rat liver microsomes. The result showed that epiberberine may have phase I and phase II metabolism in the rat liver and two metabolites in phase I and three metabolites in phase II are identified in the temperature incubation system of in vitro liver microsomes. Epiberberine showed significant inhibition on CYP2D6 with IC50 value of 35.22 μmol L(-1), but had no obvious inhibiting effect on the activities of CYP3A4, CYP1A2, CYP2E1 and CYP2C9. The results indicated that epiberberine may be caused drug interactions based on CYP2D6 enzyme. This study aims to provide a reliable experimental basis for its further research and development of epiberberine.
Animals ; Berberine ; analogs & derivatives ; chemistry ; metabolism ; Chromatography, High Pressure Liquid ; Cytochrome P-450 CYP2D6 ; metabolism ; Cytochrome P-450 CYP2D6 Inhibitors ; chemistry ; metabolism ; Drugs, Chinese Herbal ; chemistry ; metabolism ; Male ; Microsomes, Liver ; drug effects ; enzymology ; metabolism ; Molecular Structure ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry

OBJECTIVETo observe different degrees of intra-abdominal pressure and different duration on the intestinal permeability and endotoxin/bacteria translocation in rabbit model, so as to explore the mechanism of the development of abdominal compartment syndrome (ACS) and MODS.

METHODSRabbit model of intra-abdominal hypertension was established by injection of gaseous nitrogen into the peritoneal cavity. Thirty-nine New Zealand white rabbits were employed in the study. The change in intestinal permeability was determined by fluorescein isothiocyanate dextran (FITC-D) and two kinds of molecular probes of type II horseradish peroxidase (HRP-II). The effects of intra-abdominal hypertension on the endotoxin/bacteria translocation were also detected.

RESULTSThe contents of FITC-D and HRP-II in portal veins increased evidently (P < 0.01) when intra-abdominal pressure (IAP) was higher than 20 mmHg. The endotoxin (ET) content in portal vein in rabbits with IAP of 10 mmHg for 1, 2 and 4 hours exhibited no difference compared with that in normal control, while the ET content increased obviously after 1 hour with IAP of 20 mmHg and increased thereafter along with the prolongation of IAP, and increase in pressure. The bacterial translocation rates were 33.3%, 66.7% and 100% when IAP was maintained at 20 mmHg for 1, 2 and 4 hours, respectively, and there was evidence of bacterial translocation to the liver. The rate of bacterial translocation to intestinal mesenteric lymph nodes was 100% when IAP was 30 mmHg for 1 and 2 hours. There was no bacterial translocation to the spleen in all experimental rabbits.

CONCLUSIONIntestinal mucosal permeability increased significantly with increased endotoxin content in portal vein when IAP was higher than 20 mmHg. At the sane time, the bacteria could be translocate to intestinal mesenteric lymph nodes and liver, which might be constitute one of the important factors leading to the development of ACS and MODS.

Abdomen ; microbiology ; physiopathology ; Animals ; Bacterial Translocation ; Colony Count, Microbial ; Compartment Syndromes ; etiology ; Endotoxins ; blood ; Female ; Intestines ; physiopathology ; Male ; Multiple Organ Failure ; etiology ; Permeability ; Rabbits

BACKGROUNDTumor necrosis factor alpha (TNF-α) is important in promoting relative adrenal insufficiency (RAI) due to systemic inflammatory response syndrome (SIRS). We identified the TNF-α receptor involved in the inhibition of adrenal corticotrophin (ACTH)-stimulated hydrocortisone release by studying the expression of TNF-α receptors in adrenal cortex Y1 cells and the effect of downregulating TNF receptors on ACTH-stimulated hydrocortisone release.

METHODSWe used real-time PCR and immunocytochemistry to evaluate the expression of TNF receptors on Y1 cells. TNF-receptor 1 (TNF-R1) DNA fragments corresponding to the short hairpin RNA (shRNA)-sequences were synthesized and cloned into pcDNA(TM) 6.2-GW/EmGFP expression vector. Knockdown efficiency of TNF-R1 expression was evaluated in miRNA transfected and mock-miRNA transfected Y1 cells by quantitative real-time PCR (Q-PCR). Hydro-cortisone expression levels were determined in TNF-R1-knockdown and control Y1 cells treated with TNF-α and ACTH.

RESULTSMouse adrenal cortex Y1 cells were positive for type I TNF-R1, but not type II TNF-receptor (TNF-R2). Blocking TNF-R1 expression resulted in loss of TNF-α-mediated inhibition of ACTH-stimulated hydrocortisone expression, suggesting a role for the TNF-R1 related signaling pathway in ACTH-stimulated hydrocortisone synthesis.

CONCLUSIONThe inhibitory effect of TNF-α on ACTH-stimulated hydrocortisone synthesis was mediated via TNF-R1 in adrenal cortex.

Adrenal Cortex ; cytology ; drug effects ; metabolism ; Animals ; Cell Line ; Hydrocortisone ; metabolism ; Immunohistochemistry ; Mice ; Real-Time Polymerase Chain Reaction ; Receptors, Tumor Necrosis Factor, Type I ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Necrosis Factor-alpha ; pharmacology

Objective To provide updated recommendations for further adjustment of iodine deficiency disorders control strategy by evaluating awareness of the dangers of iodine deficiency disorders and their prevention and control measures among local residents and the willingness of choosing iodized or non-iodized salt in urban communities of Jiangsu province.Methods Trained health service workers did questionnaire investigation on iodine deficiency disorders prevention knowledge by calling computer-randomly selected family telephone numbers in every two blocks of provincial capital (Nanjing) and coastal city (Nantong) in Jiangsu province in 2010.The survey topics covered include:when supplied with both non-iodized and iodized salt,what kind of salt the local residents chose and the reasons; residents awareness of the dangers of iodine deficiency disorders and their prevention and control measures; resident's personal information(including only occupational category and age group).Key fields were rechecked from 10％ randomly selected valid questionnaires.Results Four hundred and fifty-five valid questionnaires were obtained by calling 2021 randomized telephone numbers in Nanjing and Nantong cities.After 10.1％(46/455) of the valid questionnaires were rechecked,the total coincidence rate was 87.0％(40/46).In the two cities,there were 73.2％ (333/455) of those 455 respondents saying they would choose iodized salt when iodized and non-iodized salt were supplied at the same time.There were 69.0％(314/455) of respondents had heard of iodine deficiency disorders,of which 79.6％(250/314) of them were aware of the dangers of iodine deficiency is endemic goiter,and 10.8％ (34/314) of them knew that iodine deficiency can cause varying degrees of mental damage.With respect to useful preventive methods of iodine deficiency disorders,there were 69.4％ (218/314) respondents know that eating iodized salt,and 41.1％ (129/314) respondents know that taking laver or kelp would prevent iodine deficiency disorders.Conclusions Remarkable achievements on iodine deficiency disorders health promotion and health education have been gained in urban communities of both the provincial capital and the coastal city Jiangsu province.However,some of the people are still not very clear why we should eat iodized salt.Salt industry in the current management system,is still premature to provide iodized salt and non-iodized salt simultaneously now.

OBJECTIVETo investigate the drug resistance of flavobacterium and its ability to produce BLA (beta-lactamases) and ESBLs (Extended-spectrum beta-lactamases).

METHODSThe production of BLA and ESBLs from 6 clinical isolated flavobacterium strains was determined by nitrocefin disc test and double-disc synergy method, respectively. The antibiotic susceptibilities of the strains were determined by Kirby-Bauer disc diffusion test and the agar dilution method and the MIC was assessed.

RESULTSAll the six flavobacteria were BLA-producing strains and more than 80% of them were ESBLs-producing, and they were highly resistant to beta-lactamase antibiotics (MIC 32 - 256 mg/L), but susceptible to fluoroquinolones and cephalosporin with beta-lactamase inhibitors (MIC 0.125 - 8 mg/L).

CONCLUSIONMost of the flavobacteria in nosocomial infections were beta-lactamase-producing and were highly resistant to beta-lactamase antibiotics. Fluoroquinolones and beta-lactamase antibiotics with lactamase inhibitors should be the first choice for the management of infection caused by flavobacterium.

Anti-Bacterial Agents ; pharmacology ; Drug Resistance, Bacterial ; drug effects ; Flavobacterium ; drug effects ; enzymology ; Humans ; Membrane Proteins ; analysis ; metabolism ; Microbial Sensitivity Tests ; Ribosomal Proteins ; analysis ; metabolism

This study was aimed to investigate the effect of baicalin on proliferation and apoptosis of HL-60 cells and its mechanism. Cell proliferation was assayed by using Cell Counting Kit-8. The morphological changes of HL-60 cells were examined by light microscopy and nucleolus morphological changes were observed by fluorescent microscopy after Hoechst 33342 staining. The early cell apoptosis was detected by using flow cytometry with Annexin V-FITC/PI double staining. The expression of caspase-3, caspase-9, Bcl-2 and Bax mRNA was detected by RT-PCR and Western blot assay was carried out to examine Bax, Bcl-2, caspase-8 and cleaved caspase-3 expression. The results showed that Baicalin inhibited the proliferation of HL-60 cells in a time- and concentration-dependent manner. HL-60 cells exhibited typical morphological features (for example, cell shrinkage, membrane blebbing and formation of apoptotic bodies). Cell apoptosis in early stage could be detected, the expression of caspase-3, caspase-9 and Bax mRNA was obviously up-regulated, while the Bcl-2 expression down-regulated, and accordingly Bcl-2/Bax ratio decreased. Such results were consistent with the expression of these proteins. In addition, the expression of cleaved caspase-8 protein was induced significantly after treated with baicalin. It is concluded that baicalin can significantly inhibit the proliferation of HL-60 cells and induce the apoptosis of HL-60 cells, which may occur through decreasing Bcl-2/Bax ratio by intrinsic pathway and through extrinsic pathway. It suggests that baicalin may be a promising drug for the therapy of acute myeloid leukemia.
Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Cell Proliferation ; drug effects ; Flavonoids ; pharmacology ; HL-60 Cells ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism