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Objective To evaluate the performance of Xpert C. difficile multiplex real-time PCR assay for diagnosis of Clostridium difficile infections in Chinese hospital settings. Methods This study was performed in Huashan Hospital, Ruijin Hospital, Beijing Hospital, Nanfang Hospital and Sir Run Run Shaw Hospital using a standard study protocol. Unique unformed stools from patients with acute hospital-acquired diarrhea were simultaneously analyzed by toxigenic anaerobic cultures and the Xpert C. difficile assay. All specimens displaying discordant results between the Xpert assay and toxigenic culture were sent for Sanger tcdB gene sequencing. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), total concordance rate, and 95％ confidence interval (CI) were calculated before and after resolution of discordant results using SAS 9.3. Results A total of 745 stool specimens were collected and 46 were excluded due to failure of C. difficile recovery. The remaining 699 specimens were included. Compared to the results of toxigenic culture, the sensitivity, specificity, PPV, and NPV of Xpert C. difficile assay were 94.1％ (144/153)(95％CI:89.1％-97.3％), 93.2％ (509/546)(95％CI:96.7％-99.2％), 79.6％ (144/181)(95％CI:72.9％-85.2％)and 98.3％ (509 / 518) (95％CI: 96.7％-99.2％), respectively. Both methods had a Kappa of 0.819. Xpert C. difficile assay showed sensitivity of 98.4％(62/63) (95％ CI: 90.3％-99.9％) and specificity of 93.2％(509/546) (95％ CI: 90.8％-95.2％) for toxin A-negative toxin B-positive strains. After the discordant results resolved by tcdB gene sequencing, PCR assay provided better performance with high sensitivity, specificity, positive predictive value, and negative predictive value [98.8％ (171 / 173), 98.1％ (516 / 526), 94.5％ (171/181) and 99.6％ (516/518), respectively]. Conclusions Compared to the results of toxigenic culture, the sensitivity, specificity and NPV of Xpert C. difficile assay were 94.1％ (144/153) and 93.2％(509/546), respectively. With the results available within 1 h, Xpert C. difficile assay provides prompt and precise laboratory diagnosis in Chinese clinical settings.
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Objective To compare the short-term curative effects of prucalopride and mosapride respectively combined with low dose polyethylene glycol in treating elderly refractory functional constipation.Methods Ninety patients with elderly refractory functional constipation in the outpatient department of our hospital from May 2014 to February 2016 were retrospectively analyzed and divided into two groups randomly,45 cases in each group.the prucalopride group:Prucalopride Succinate Tablets,2mg,4 times daily;the mosapride group:Mosapride Citrate Capsules,5mg,3 times daily.Polyethylene Glycol Electrolytes Powder(PEG) was also used in the two groups,13.125g,twice daily.The course of treatment was 4 weeks.The first defecating time and defecation difficulty relief time,average weekly spontaneous complete bowel movements (SCBM),defecating difficulty,stool character,adverse reactions and change of life quality were observed in the two groups.Results The treatment effective rate of slow transit constipation(STC) and defecatory disorder had the statistical difference between the two groups (P0.05).Compared with the mosapride group,the first defecating time and defecation difficulty relief time in the prucalopride group were shorter with statistical difference (P0.05).The total effective rate had statistical difference between the two groups(P<0.05).The incidence rate of adverse reactions had no statistical difference between the two groups(17.78% vs.15.56%,P<0.05).The total average score of PAC-QOL after treatment in the two groups were both decreased,moreover the decrease in the prucalopride group was more obvious;the difference between the two groups had statistical significance(P<0.05).Conclusion Prucalopride +PEG take effect faster in the treatment of elderly refractory functional constipation and has the advantages in the aspects of overall curative effect and life quality improvement,which is specially suitable for STC and defecatory disorder type.
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Objective To investigate the role of spo0A gene in growth and sporulation of Clostridium difficile clinical isolates. Methods ClosTron gene knock-out system was used to knock out the spo0A gene of C. difficile strain C25. Bacterial growth curve was plotted by measuring D600 with spectrophotometer in different phases of bacterial growth. Malachite green staining technique was used to count the number of vegetative cells and spores under optical microscope. The sporulation rate was calculated. Results The spo0A mutant and its C25 parental strain showed similar patterns of growth. However, after knock-out of spo0A gene, an asporogenous phenotype was built, while the parental strain could produce spores as usual.Conclusions The spo0A gene plays a key role in sporulation but not growth of C. difficile strain.
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Objective To evaluate the in vitro antibacterial activity of fusidic acid against P.acnes.Methods Fifty strains of P.acnes were clinically isolated from Huashan Hospital,Fudan University from March to September 2013.The minimum inhibitory concentrations (MICs) of several antibacterial agents including fusidic acid against these P.aches isolates were determined by using the agar dilution method according to the recommendations of Clinical and Laboratory Standards Institute (CLSI).Data were analyzed using the WHONET 5.4 software.Results Among the 50 P.acnes isolates,90％ were sensitive to fusidic acid,90％ to moxifloxacin,54％ to clindamycin,46％ to erythromycin,but 100％ were resistant to metronidazole.The minimum concentration required to inhibit the growth of 50％ organisms (MIC50) and 90％ organisms (MIC90) were 0.5 and 1.0 mg/L respectively for fusidic acid,whereas clindamycin and erythromycin both showed higher MIC90 values (＞ 128 mg/L).At the concentration of 128 mg/L,clindamycin inhibited the growth of 70％ of the P.acnes isolates,and erythromycin inhibited the growth of 48％ of them,while the growth of all the isolates was inhibited by fusidic acid at 2 mg/L.Conclusion Fusidic acid exhibits excellent antibacterial activity against clinical isolates of P.acnes in vitro.
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Objective To determine the in vitro antibacterial activity of levornidazole against 375 anaerobic isolates.Methods Agar dilution method was used to determine the minimum inhibitory concentrations (MICs)of levornidazole,3 comparators (metronidazole,ornidazole and dextrornidazole)against 375 anaerobic isolates.Results For anaerobic gram-negative and gram-positive bacilli,and anaerobic gram-positive cocci,levornidazole displayed activity similar to or slightly higher than that of met-ronidazole,ornidazole and dextrornidazole.Levornidazole showed good activity against B.fragilis,Bacteroides thetaiotaomi-cron ,Clostridium difficile ,Clostridium perfringens ,and Peptostreptococcus magnus .The MIC90 value of levornidazole a-gainst the above-mentioned anaerobes was 0.5,1,0.25,2 and 1 mg/L,respectively.However,levornidazole and the compa-rators had poor antibacterial activity against Veillonella spp.among anaerobic gram-negative cocci.Conclusions The in vitro anti-anaerobic activity of levornidazole is similar to or slightly higher than that of metronidazole, ornidazole and dex-trornidazole.Levornidazole has good activity against both gram-negative and gram-positive anaerobic bacilli,and gram-positive anaerobic cocci,suggesting its promising clinical use.
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Objective To explore a method for establishing the hamster model of Clostridium difficile-associated diarrhea (CDAD)and the indicators for its evaluation.Methods Clindamycin was administered to hamsters subcutaneously (day 1),and 24 h later infected with C.difficile clinical isolates KH1 (ribotype 027,106-108 CFU/mL)or SH9 (ribotype 001 ,108-1010 CFU/mL)by gavage.Animals were observed for CDAD symptoms such as diarrhea,weight loss and death.At the end of ob-servation period (day 7 or death),the cecum was collected from each animal for histological evaluation of inflammation.Results Following a single dose of 100 mg/kg clindamycin subcutaneously,all the animals challenged with KH1 (108 CFU/mL)devel-oped diarrhea and then died within 5 days.All the hamsters challenged with SH9 (1010 CFU/mL)developed diarrhea as well but only 66.7% died at the end of observation period.Among other groups,only one or none developed diarrhea and then died. The symptoms of hamsters with diarrhea included loose stool,wet tail and weight loss.On histological examination,conges-tion,hemorrhage and neutrophil infiltration of the mucosa were observed in the hamsters died of CDAD.Conclusions We have successfully established a hamster CDAD model that allows for future investigations.
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Background:Psychological factor plays an important role in the pathogenesis of functional gastrointestinal disorders. Depression,anxiety may share the pathogenesis with irritable bowel syndrome( IBS),therefore,more and more attention has been paid to anti-depression and anti-anxiety in the treatment of IBS. Aims:To evaluate the effect of addition of flupentixol and melitracen tablets on IBS with depression and anxiety. Methods:Sixty IBS patients with depression and anxiety were randomly divided into control group and treatment group. Patients in control group were given live combined Bacillus subtilis and Enterococcus faecium enteric-coated capsules and trimebutine maleate,while paitents in treatment group were received flupentixol and melitracen tablets in addition to the above-mentioned medicine. Treatment course was 8 weeks. Hamilton depression scale,Hamilton anxiety scale and symptom score were evaluated before and after treatment. Results:Compared with control group,depression and anxiety in treatment group were obviously improved(P<0. 05);the relief rates of abdominal pain and diarrhea were significantly increased( P <0. 05 );and the total symptom score were significantly decreased(P<0. 05);the total effective rate was significantly increased(93. 3% vs. 76. 7%,P<0. 05). However,no significant difference in incidence of adverse reaction was found between the two groups. Conclusions:Additon of flupentixol and melitracen tablets can significantly improve the curative effect on IBS with depression and anxiety.
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0.05).CONCLUSION:The domestic tropisetron hydrochloride is equivalent to the imported one in terms of the efficacy and safety in the treatment of cisplatin-induced nausea and vomiting.
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Objective Taking CNE-2Z multicellular spheroids (MCSs) as the simulation of solid tumors, to investigate the role of SAPK/JNK signaling pathway in multicellular resistance to radiotherapy for human nasopharyngeal carcinoma (NPC). Methods Human NPC cell line CNE-2Z were cultured into multicellular spheroids by using liquid overlay technique, then divided into control MCSs, irradiated MCSs (average dose in one minute: 2 Gy), sp-600125(a specific inhibitor for SAPK/JNK signaling pathway)+irradiated MCSs, sp-600125+MCSs. Western blotting was employed to analyze the activity of SAPK/JNK signaling pathway in MCSs, and the expression of Caspase-3 protein before and after sp-600125 treatment; X-ray induced cell apoptotisis in MCSs before and after sp-600125 treatment was detected by TUNEL. Results The level of SAPK/JNK phosphorylation in MCSs was a dynamic course after radiation, and the phosphorylation peaked at 2 h after irradiation; The apoptotic rate of MCSs (P
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Objectives To explore the effect of NF-?B signaling pathway on multicellular resistance(MCR) to 5-Fu in colon carcinoma cells.Methods The multicellular spheroids(MCS) of colon carcinoma HT-29 cell line were cultured by using liquid overlay technique,and the monolayer cells by using routine procedure.The morphology of MCS was observed with light microscope and scanning electronmicroscope(SEM).The expression of proliferating cell nuclear antigen(PCNA) was determined by immunohistochemistry.24 hours after adding 50?g/ml SN50 to spheroids cells,the 50% inhibiting concentration(IC50) for 5-Fu was determined by using radial outgrowth assay.The activity of NF-?B was measured by using electrophoretic mobility shift assay(EMSA) and the specific bands were observed.Apoptosis of spheroids cells,which were pretreated with 5?g/ml 5-Fu for 24 hours,was identified by TUNEL,the apoptosis related proteins caspase-3,Bcl-2 and Bax were detected by Western blotting and analyzed by semi-quantitative method.Results A necrotic core formed in the centre of MCS,and the peripheral cells were actively proliferating.Compared with the monolayer cultured cells,the activity of NF-?B was increased in MCS.When the activity of NF-?B in spheroid cells was inhibited by SN50,5-Fu readily induced cells to apoptosis(15.75%?1.02% vs 8.71%?0.73%,P
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Objective To investigate the changes in cells on cell cycle,cell apoptosis and susceptibility to chemotherapeutic drugs of lung adenocarcinoma A549 cells under two dimensional (2D system) and three dimensional (3D system) culture conditions. Methods The three dimensional culture of A549 cells was carried out using rotatable culture bottle and desk-top water bath shake. Flow cytometer,in situ cell apoptosis detection kit,MTT and blood cell counter were employed to compare cell cycle,cell apoptosis and susceptibility to ADM of A549 cells under different culture conditions. Results The three dimensional culture system was confirmed to have been successful by the aid of observation under inverted and electron microscope. There were significant retardation of G_ 1 phase,lower cell apoptosis rate and decreased susceptibility to ADM in A549 cells in 3D system compared with those in 2D system. Conclusions There were remarkable differences in biological characteristics of A549 cells in two culture systems,indicating that A549 cells cultivated in 3D system simulated better solid tumor in vivo . The 3D system was very useful for further investigation of the behavior of solid tumor,so that anti-carcinoma chemotherapeutic drugs could be advantageously tested in vitro before clinical application.
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OBJECTIVE To investigate the trend of resistance of antimicrobial agents among clinical isolates of Klebsiella pneumoniae 1996-2005.METHODS Agar diffusion test was used to analyze the changes in drug susceptibility of K.pneumoniae isolated from 11 hospitals in Shanghai to antimicrobial agents.RESULTS The resistance rates of K.pneumoniae to antimicrobial agents mostly increased 1996-2005.The resistance rates to piperacillin,cefazolin and cefuroxime remained at high levels(more than 50%),while which to cefotaxime(29.3-45.2%) and piperacillin/tazobactam(5.1-20.6%) were increased obviously.Carbapenems were the most active agents against the K.pneumoniae isolates.The resistance rates to ciprofloxacin increased from 14.4% to 35.8%.The detectable rate of extended-spectrum beta-lactamases-producing strains in K.pneumoniae increased from 30.1% to 42.0%.CONCLUSIONS The trend of resistance of K.pneumoniae to commonly used antimicrobials was upward 1996-2005.