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OBJECTIVE: To establish GC-MS fingerprint of volatile oil from Citrus aurantium. METHODS: GC-MS method was adopted. The determination was performed on RTX-5MS capillary column with injector temperature of 250 ℃, high pure helium as carrier gas(≥99. 999％), flow rate of 1. 0 mL/min, split ratio of 10:1,and sample size of 1 μL (temperature programming). Mass spectrum condition included electron bombardment ion source, ion source temperature of 230 ℃, detector temperature of 250 ℃, 3 min solvent delay, scanning range of m/z 35-550. GC-MS chromatograms of 21 batches of volatile oil samples were determined using Laurene as reference. The similarity of them was evaluated by using TCM Chromatographic Fingerprint Similarity Evaluation System (2004 A edition), and common peak was determined. The components of common peak were determined by LC Solution 2 mass database (NIST05. LIB and NIST05s. LIB). Relative content of common peak was determined with area normalization. RESULTS; There were 20 common peaks in GC-MS chromatograms of 21 batches of volatile oil samples, and the similarity was higher than 0. 90. After validation, GC-MS chromatograms of 21 batches of volatile oil samples were in good agreement with control fingerprint. The main constituents of the volatile oil of C. aurantium were Limonene, Terpinene, Laurene and D-Cadinene. CONCLUSIONS: Established fingerprint can provide reference for identification and quality evaluation of volatile oil of C. aurantium.
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Objective To compare clinical Pseudomonas aeruginosa different sources of β-lactamase-resistant phenotype differences,as to provide theoretical basis for guiding clinical rational use of antibiotics.Methods Isolation of 478 strains of Pseudomonas aeruginosa were collected from clinical specimens in the First Affiliated Hospital of Xi'an Jiaotong University from January to December 2015,by VITEK 2 Compact bacteria identification and drug sensitivity analysis of advanced expert system for β-lactamase-resistant phenotype,statistical analysis of drug resistance phenotype and antibiotic resistance.Results 478 strains of Pseudomonas aeruginosa were mainly composed of phenotype 5 and phenotype 3.Sputum,drainage fluid and bile duct bile specimens of Pseudomonas aeruginosa were based on phenotype 5,accounted for 31.08％,34.71％ and 38.46％.Multiple comparison x2 were 3.893,4.071 and 5.595,There was no statistical difference between groups compare significance (P＞0.05).Urine,secretions and whole blood samples of Pseudomonas aeruginosa were phenotype 3,accounted for 34.88 ％,27.78 ％,45.45 ％;Multiple comparison x2 were 6.654,9.956 and 9.852.There was no statistical difference between groups compare significance (P＞0.05).Sputum,drainage of liquid,bile duct bile and urine,secretion,whole blood specimens respectively source of Pseudomonas aeruginosa resistant phenotype distribution of two comparative difference was statistically significant (x2 =15.056～22.050,P＜0.05).Comparing the resistance of different β-lactamase-resistant phenotypes in Pseudomonas aeruginosa isolates from different sources:the sputum specimen source in imipenem,meropenem,piperacillin and piperacillin/tazobactam had significant difference (x2 =22.225～39.025,P＜0.05).There was statistical significance in department of hepatobiliary surgery only ceftazidime and meropenem differences (x2 =21.890～22.872,P＜0.05).Conclusion The phenotypic analysis of β-lactamase-resistant phenotypes of Pseudomonas aeruginosa from different specimens was different,which provided a theoretical basis for guiding the clinical application of antibiotics and the control of nosocomial infection of Pseudomonas aeruginosa.
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Objective To establish a double-antibody sandwich ELISA for the rapid detection of shiga toxin typeⅡ ( StxⅡ) in shiga toxin-producing Escherichia coli ( STEC) infection. Methods A pool of murine hybridomas was used to screen out the optimal antibody pair for the establishment of double-anti-body sandwich ELISA. The established ELISA system was used to detect StxⅡin the culture supernatants of 16 clinical strains of STEC. Specificity and sensitivity of the established ELISA system were also evaluated. Results Two antibodies, S2D8 and S2C6, were successfully screened out, based on which the double-anti-body sandwich ELISA was set up. StxⅡand its variants rather than StxⅠwas detected in the culture super-natants of STEC with a lowest detection limit of 4 ng/ml. Its performance was consistent with that of commer-cial colloidal gold test kit, indicating the characteristics of good specificity and sensitivity. Conclusion The S2D8/S2C6-based ELISA laid a foundation for researches which designates the shiga toxin as a potential can-didate on the diagnosis and therapy of STEC infection.
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Objective To explore the effects of the different doses of propofol for preventing postoperative agitation in chil-dren.Methods 60 children cases undergoing elective indirect inguinal hernia hernioplasty were selected in the Affiliated Hospital of Traditional Chinese Medicine,Luzhou Medical College from June 2011 to April 2011 and randomly divided into the group Ⅰ,Ⅱ andⅢ.The three groups were performed the general anesthesia with sevoflurane and postoperatively given 0.90% sodium chloride in-jection 0.10 mL/kg by intravenous injection,propofol 1.00 mg/kg by once intravenous injection and propofol 1.00 mg/kg by twice intravenous injection,respectively.The occurrence rate of postoperative agitation within 30 min was compared among 3 groups.The anesthesia recovery agitation score,improved Aldrete score,awakening time and time out of the operation room were also compared. Results The occurrence rates of agitation within postoperative 30 min in the group Ⅰ,Ⅱ and Ⅲ were 65.00%,65.00% and 15.00% respectively,the difference among three groups was statistically significant (P 0.05).Conclusion Postoperative twice intravenous injection of propofol 1.00 mg/kg has obvious effect and good safety for preventing the postoperative agitation in children,which has important clinical reference value.
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OBJECTIVE:To focus on the situation of clinical pharmacists particpating in the clinical drug treatment consulta-tion in China. METHODS:With the keywords of“clinical pharmacists”,“consultation”and others,retrieved from China Hospital Knowledge Database (CHKD),Wanfang Database and China BioMedical Literature Service System (SinoMed),the situation of clinical pharmacists particpating in the clinical drug treatment consultation in China was concluded and summarized. RESULTS:A total of 186 literatures were included,invloving 114 effective literatures. 2 606 cases were clinical pharmacists particpating in the clinical drug treatment consultation,2 257 cases were all or part of adoption of clinical pharmacists’suggestions,the execution lev-el of clinicians for clinical pharmacists’consultation suggestions reached 93.02%. After carrying out the clinical pharmacists’con-sultation suggestions,patients’condition improved or cured reached 2 290 cases,with the effective rate of 94.47%. CONCLU-SIONS:Clinical pharmacists participating in the clinical consultation has certain function,however,the working method and mode need to be further explored and standarized.
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Laser-induced breakdown spectroscopy ( LIBS) was used to detect the compositions of soil in the air, and the quantitative analysis model with genetic algorithm-partial least squares ( GA-PLS ) was established. A total of fifty-eight soil samples were split into calibration, monitoring and prediction sets. Eleven soil compositions including Mn, Cr, Cu, Pb, Ba, Al2 O3 , CaO, Fe2 O3 , MgO, Na2 O, and K2 O were quantitatively analyzed. The results demonstrated that, as a pretreatment method for optimizing the selection of spectral lines, GA could be effectively used to reduce the number of spectral lines for use in building PLS model, and hence simplify the quantitative analysis model. More importantly, for most of the soil compositions, GA-PLS could significantly improve the prediction ability compared with the conventional PLS model. Take Mn as an example, the root-mean-square error of prediction ( RMSEP ) was decreased from 0. 0215% to 0 . 0167%, and the mean percent prediction error ( MPE ) was decreased from 8 . 10% to 5 . 20%. The research provides an approach for further improving the accuracy of LIBS quantitative analysis in soil.
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Objective The purpose of the study was to investigate the changes and clinical significance of matrix metalloproteinase (MMP)-1,MMP-3,tissue inhibitor of metalloproteinase (TIMP),interleukin (IL)-1,tumor necrosis factor (TNF)-α and transforming growth factor(TGF)-β in juvenile idiopathic arthritis (JIA).Methods Thirty-two JIA subjects and 28 controls (traumatic arthritis patients) were included into this study.The MMP-1,MMP-3,TIMP,IL-1,TNF-α and TGF-β level in the serum and synovia were assessed by ELISA.The WBC count,the level of CRP,ESR,RF were also detected.Independent t-test and Pearson's analysis were adopted for data analysis.Results ① The level of MMP-1,MMP-3,IL-1 and TNF-α in the serum was (158±67) ng/ml,(212±89) ng/ml,(39±19) pg/ml,(26±10) pg/ml respectively,which was significantly higher than that of the control group (all P＜0.05) ; the ratio of MMP-3/TIMP-1 (0.86±0.32) was higher in the study group than that of the control group (P＜0.05),while the value of TIMP-1,TGF-β was (248±88),(17±9) ng/ml respectively,which was lower than that of the control group (P＜0.05).The value of seral MMP-3,MMP-3/TIMP-1 was positively correlated with that of WBC,CRP,ESR in the JIA group (all P＜0.05).② The value of MMP-1,MMP-3,IL-1 and TNF-α in the synovia was (216±78) ng/nl,(766±291)ng/ml,(56±21) pg/ml,(36±14) pg/ml respectively,which was higher than that of the control group (all P＜0.05); the ratio of MMP-3/TlMP-1 (2.68±0.89) was higher than that of the control group (P＜0.05),while the value of TIMP-1,TGF-β was (286±88) ng/ml,(12±4) ng/ml respectively,which was lower than that of the control group (all P＜0.05).③ The value of MMP-1,MMP-3,TIMP-1,IL-1 and TNF-α in the synovia was higher than that in the serum (all P＜0.05),while the value of TGF-β was lower than that in the serum (P＜0.05).Conclusion The value of MMP-1,MMP-3,IL-1 and TNF-α increases both in the serum and synovia,while the value of TIMP-1 decreases.The value of TGF-β decreases,which may have protective effect on JIA.The ratio of MMP-3/TIMP-1 in the serum is positively correlated to inflammation parameters,which may be used to judge the activity of illness in JIA.
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Shiga toxin 2 (Stx2) toxoid produced by formaldehyde treatment of purified toxin was used to immunize BALB/c mice for monoclonal antibody (MAb) production.The neutralizing activities of positive clones against Stx2 were screened by in vitro cytotoxicity assay.The isotype and specificity of resultant clone was determined,and its efficacy to neutralize the activity of purified Stx2 was evaluated by in vitro and in vivo toxicity model.Lastly,its spectrum of activity against Stx2 variants was also accessed by mouse toxicity model.It was demonstrated that one of the 12 positive MAb clones against Stx2,designating $2C4 had neutralizing activity.S2C4 belongs to the immunoglobulin G1 subclass and has a K light chain,and it reacts with the A subunit of Stx2 and does not bind to Stx2 B subunit or to Stx1.S2CA could efficiently neutralize the cytotoxicity of Stx2 to Veto cells and mice.It also protected mice against lethal doses of Stx2 variants challenge including Stx2c and Stx2vha.S2C4 is a promising candidate molecule in preventing the progression of hemolytic-uremic syndrome (HUS) mediated mainly by Stx2 in Stx-producing Escherichia coli(STEC)infection.
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Objective To purify Shiga toxin Ⅱ (STX Ⅱ) of enterohaemorrhagic Escherichia coli (EHEC) O157: H7 by affinity chromatography, and characterize its biological function. Methods The immno-affinity chromatography column was prepared by STX Ⅱ A subunit-specific antibody S1D8 coupling to Sepharose 4B matrix. The purity and specificity of STX Ⅱ molecule secreted by EHEC O157:H7 were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot, respectively. The purified toxin was serially diluted and the toxic activities to Vero cell line and mice were observed. The 50% cytotoxic dose (CD50) for Vero cell line and 100% lethal dose (LD100) for mice were calculated. The protection effect of anti-STX Ⅱ polysera to the mice against the purified toxin challenge was also observed. Results STX Ⅱ was successfully purified from culture supernatant of EHEC O157:H7 using affinity chromatography scheme. The relative molecular weights of STX Ⅱ A and B subunits were 32 000 and 7 500 confirmed by SDS-PAGE, respectively. The purified toxin could react with monoclonal antibodies against STX Ⅱ A and B subunits, respectively.The toxin was cytotoxic to Vero cell with CD50 of 20 ng/L and lethal to mice with LD100 of 5 ng.The toxin could be neutralized by anti-STX Ⅱ polysera in vivo. Conclusion STX Ⅱ is successfully purified and its toxic effects are confirmed in both cell line and mouse model.