The paper reports the establishment of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) for simultaneous analysis of 11 opiates in hair samples, and the study of presence of opiates in the hair of active heroin addicts. About 20 mg of decontaminated and pulverized hair sample was hydrolyzed with buffer solution for 30 min, in the presence of morphine-d3 and acetylmorphine-d6 used as internal standards, and then extracted with the mixture of dichlormethane and isopropanol, separated by the Allure PFP propyl column with a mobile phase consisting of acetonitrile and 20 mmol L(-1) ammonium acetate buffer, and then analyzed by LC-MS/MS. Multiple reaction monitoring (MRM) mode was used to analyze 11 opiates. Eleven opiates showed a fairly good linearity over the corresponding range (r > 0.996 0). The detection limits were less than 0.05 ng mg(-1). The recoveries were between 47.2% and 110%, and the deviations of intra- and inter-day precision were less than 14%. Heroin, acetylmorphine, morphine, codeine, acetylcodeine and hydrocodone were detected in hair samples of 21 herion addicts. The developed method shows high sensitivity and selectivity, and is suitable for the simultaneous analysis of 11 opiates in hair samples and identify legal and illegal use of opiates.

Objective To discuss the best resistance factors of paclitaxel(Taxinol)on benign biliary scar fibrosis,in order to provide an effective basis for clinical prevention and treatment of benign biliary scar fibrosis.Methods Human bile duct epithelial cells were cultured in vitro,the prepared PTX at 0.001 uM,0.005 uM,0.1 uM,0.5 uMand 1 uMconcentration were separately added into cells for 48 h.The half inhibitory rate of BEC (IC50) were determined by MTT and the optimal concentration were confirmed.Human bile duct epithelial cells were cultured in 0 h,24 h,48 h and 72 h,the inhibitory rate of BEC at 100 nM,250 nM,and 500 nM PTX-Chitosan Sustained release membranes and the optimal concentration of PTX were determined by MTT and the optimal concentration of PTX-SRM were obtained.Human bile duct epithelial cells were cultured for 48 h and 72 h,the mRNA and protein expression ofα-SMA,E-cadherin,Vimentin were detected by Western Blot and Real-time PCR methods.Results The optimum resistance concentration of PTX to benign biliary scar was 250 nM.PTX and PTX-SRM could effectively inhibit the proliferation and transformation of BEC,and the best effective treatments to resist benign biliary scar fibrosis were low and middle concentrations of PTX-SRM,the best drug loading were 100 nMand 250 nM.The inhibition duration of PTX-SRMon BEC was longer than PTX alone(P<0.05).Conclusion The inhibition of PTX-SRMon BEC proliferation and transformation is better than the single drug of PTX,which provides a new scientific and feasible method for clinical prevention and treatment of benign biliary scar.

Objective To explore the effect of Paclitaxel-Chitosan Sustain film on growth, apoptosis and cell cycle of biliary fibroblasts cells. Methods Human biliary fibroblasts cells were cultured and treated with PTX-CSF and naked PTX,separately, untreated cells as blank control. The experiment was divided into five groups:untreated group, simple PTX-treated group (250nM) and low, medium and high chitosan sustained-release film PTX-treated group (100 nM, 250 nM, 500 nM). The proliferations of cells were determined by MTT assay. The apoptosis and cell cycle of cells were detected by FCM. Results The proliferation of biliary fibroblasts cells was inhibited by PTX-CSF with time-dependent and dose-dependent, and the inhibiting effect was more obvious than naked PTX treatment as the time went on. Meanwhile, PTX-CSF could inhibit the magration of bile duct fibroblasts induced by TGF-β1,and had longer effect than naked PTX. After 72 h, the apoptosis rate of cells treated with PTX-CSF was significantly higher than cells treated with naked PTX or untreated cells(P<0.05), the difference between naked PTX or untreated cells was not significant. Compared with untreated cells, the proportion of G 2/M in cells treated with PTX or PTX-CSF were significantly increased, and the former was sinificantly higher than the latter(P<0.05). Conclusion Compared with naked PTX, PTX-CSF has strong cytotoxic effects and obviously sustained-release effect. The effective concentration can be maintain for a long time by PTX-CSF, and it could be as the novel drug delivery system to continuously inhibit proliferation of bile duct fibroblasts.

Duplicate submission of research papers have many negative effects on authors,hospitals,readers,and journals.Reasons for duplicate submission were analyzed.Through strengthening the propaganda and using information management system,duplicate submission of research papers might be eliminated.

Objective To investigate the effects of intestinal microbiota of patients with chronic constipation on the expression of serotonin transporter (SERT) and the bowel movement in mice.Methods Fecal samples of patients with slow transit constipation met Rome Ⅲ criteria and healthy normal controls were collected and made into fecal microbiota solution.Twenty specfic pathogen free (SPF) mice were divided into experiment group and control group.The mice of two groups were both pre-treated with streptomycin to establish the germ-free mice model.The mice of control group were gavaged with mixed fecal microbiota solution of healthy normal controls and the mice of experiment group were gavaged with mixed fecal microbiota solution of patients with chronic constipation.Mice were sacrificed after fed for 15 days.Defecation parameters and ink discharge time of mice were detected.The expressions of SERT mRNA and SERT protein in mice intestinal tissues were detected with real time-polymerase chain reaction and Western blotting.The 5-hydroxytryptamine (5-HT) levels of mice intestinal tissues were evaluated by enzyme-linked immunosorbent assay (ELISA) and double immunofluorescent staining.The methods of t test and Chi-square test were performed for statistical analysis.Results On the 15th day,the total number of the feces within 2 h of the experiment group and control group was 8.55±1.83 and 12.14±2.90,respectively,and the difference was statistically significant (t=3.33,P＜0.05).The weight of feces were (151.90 ± 32.42) mg and (246.72 ± 64.01) mg,respectively,and the difference was statistically significant (t=4.18,P＜0.01).The dry weight of feces were (65.52±11.76) mg and (92.93±23.07) mg,respectively,and the difference was statistically significant (t=3.37,P＜0.05).The water content of feces were (56.63 ± 3.01) ％ and (61.95 ± 3.70) ％,respectively,and the difference was statistically significant (t=3.57,P＜0.05).The defecating time of first black feces of the experiment group and control group were (83.24±11.31) min and (69.06±2.72) min,respectively,and the difference was statistically significant (t=-2.74,P＜0.05).The expressions of SERT mRNA and SERT protein levels in mice intestine tissues of the experiment group were significantly higher than those of the control group (t =2.61,-6.89;both P＜0.05).5-HT level of mice intestinal tissues of the experimental group and control group were (151.69± 10.18) ng/mL and (198.77 ± 25.99) ng/mL,respectively,and the difference was statistically significant (t=-4.13,P＜0.01).Conclusion Intestinal microbiota of patients with chronic constipation may influence the expression of SERT in the mice intestinal tissues,and then decrease the level of 5-HT,slowing the bowel movement in mice.

OBJECTIVE:To study the activity of 10 chemical parts extracted from Isatis indigotica against herpes simplex virus(HSV)in vitro and to determine the antiviral active parts.METHODS:10 crude fractions and refined portions were extracted from Isatis indigotica using solvent isolation and absorption chromatography.Their inhibitory effects against HSV-Ⅰ and HSV-Ⅱ were determined by MTT using virus inhibition ratio and therapeutic index(TI)as appraisal targets to compare the antiviral effects among all the chemical parts.RESULTS:The parts(Ⅷ,Ⅹ)with EtOH(10% and 50% in concentration)elution via macroporous resin absorption showed the most potent activities,whose inhibitory rates against HSV-Ⅰ were 53.21% and 56.28% respectively,similar the action of its positive control acyclovir(62.55%),and their TI were 4.61 and 18.62 respectively.CONCLUSION:There were significant differences in the antiviral activity among all the chemical parts extracted from Isatis indigotica,

Objective To explore the clinical value of color Doppler flow imaging(CDFI) combined with quantitative analysis technique to evaluate renal flow perflusion. Methods Twenty patients with different renal diseases matched with healthy control subjects were examined with Philips iU22 CDFI system. CDFI system and Qlab quantitative technique were used to measure renal restrictive index(RI), vascularization index(VI), flow index(FI), vascularization-flow index (VFI). The patients all received renal single-photon emission computed tomography (SPECT) examination. Results The RI of 32 kidney with lower flow perfusion in SPECT were higher than in the contol group (P <0.01). VI,FI, VFI were lower than those in control(VI, VFI, P <0.01 ;FI, P <0.05). Conelusious The renal blood flow perfusion of patients with renal diseases are lower than that of healthy people. It has high clinical value using CDFI combined with quantitative analysis technique to evaluate the renal perfusion of the patients with renal diseases.

Objective To evalute the incidence and epidemiologic characteristics of Kawasaki disease (KD) in Wenzhou,China.Methods We used a questionnaire survey and reviewed the medical records and reports of all patients with KD diagnosed during the 10 year periods from January,2001 to December,2010.Results We studied 827 inpatients diagnosed with KD during the 10-years period from 2001 to 2010.There were 613 cases (74.12％) with complete KD.The ratio of male to female ratio was 2.28∶1.Age at onset ranged from 37 days to 13 years old,and the peak age group was 1 year old.The disease occurred in all of the seasons,but the peak was from April to June.The incidence of cardiovascular damage in acute KD was 34.6％,and the most common sequela was coronary artery dilatation.There were16 patients with coronary aneurysm.Fifty-three patients did not respond to immunoglobulin (6.4％),and 12 patients (1.5％) developed recurrent KD.After treatment,114 cases (13.8％) developed neutropenia.There were no deaths during hospitalization.Conclusion Patients with KD has become more and more in Wenzhou.Cardiovascular damages were similar to those in Beijing and Guangzhou,but higher than that reported in Japan.

This study aimed to examine the preparation of cationic lipid microbubble (CLM), and evaluate its physical and chemical properties and toxicity, measure the gene transfection efficiency by ultrasound triggered microbobble destruction (UTMD) in combination with CLM. The CLM was prepared by the method of the thin film hydration, and its morphology was observed under the electron microscopy at 1st, 3rd, 7th, 10th, and 14th day after preparation, respectively. The size, Zeta potential and stability of CLM were tested. The acute toxicity of CLM was assessed. The green fluorescent protein gene (EGFP) transfection efficiency was evaluated. The experiment grouping was as follows: naked plasmid group (P group), ultrasonic irradiation plus naked plasmid group (P-US group), naked plasmid plus CLM group (P-CLM group), naked plasmid plus ultrasound and CLM group (UTMD group). The expression of EGFP was detected by fluorescent microscopy and flow cytometry. The results showed that CLMs were spherical in shape, with the similar size and good distribution degree under the light and electron microscopies. The size of CLMs was varied from 250.4±88.3 to 399.0±99.8 nm and the Zeta potential of CLMs from 18.80±4.97 to 20.1±3.1 mV. The EGFP expression was the strongest in the UTMD group, followed by the P-CLM group, P-US group and P group. Flow cytometry results were consistent with those of fluorescent microscopy. The transfection efficiency was substantially increased in the P-US group, P-CLM group and UTMD group as compared with that in the P group, almost 7 times, 10 times and 30 times higher than that in the P group respectively. It is suggested that CLMs prepared by the method of thin film hydration are uniform in diameter, and proved non-toxic. UTMD combined with CLM can significantly increase the transfection efficiency of EGFP to targeted cells.

Objective To assess the value of quantitative color Doppler sonography in patients with systemic lupus erythematosus (SLE). Methods The right renal in 21 patients with SLE and 22 normal subjects were examined with Philips IU22 ultrasonic diagnostic device. Resistence index (RI) of interlobar artery and interlobular artery were measured with color Doppler sonography, vascularity index (VI), flow index (FI) and vascularization flow index (VFI) which reflecting the renal cortical blood flow were calculated with quantitated analysis software. All the parameters between two groups were compared and analyzed. Results The color signals of renal cortical blood flow in patients with SLE were inferior to those of the normal group, especially in the area near the edge of renal. No statistical difference of RI was found between the two groups (P>0.05), while VI, FI and VFI in SLE group were significantly lower than those of the normal group (VI, VFI, P<0.01, FI, P=0.01). Conclusion Color Doppler sonography combined with blood flow quantitative technique provide a helpful, sensitive and convenient tool to detecte the renal changes in SLE patients.