OBJECTIVE To establish a convenient method for detecting plasmid-mediated AmpC beta-lactamases in Enterobacteriaceae lacking chromosomal AmpC beta-lactamases and to investigqte their genotype.METHODS Fifteen isolates of Klebsiella pneumoniae were collected and the diameters of inhibitory zone of cefoxitin in combination with or without cloxacillin were measured separately by standard disc diffusion test.The M3D assay was used as control method.And the activity of AmpC beta-lactamases of all isolates was simultaneously assayed.Multiplex PCR was performed to determine the genotype of AmpC beta-lactamases.RESULTS Among 15 isolates,8 isolates were identified to have plasmid-mediated AmpC beta-lactamases.The sensitivity and specificity of AmpC beta-lactamases phenotypic confirmatory test were 100%.Eight of 15 isolates were identified to be DHA-1 beta-lactamases by multiplex PCR.CONCLUSIONS The new AmpC beta-lactamases phenotypic confirmatory test is a reliable method for detecting plasmidmediated AmpC beta-lactamases in Enterobacteriaceae lacking chromosomal AmpC beta-lactamases.DHA-1 beta-lactamases are the main genotypes of plasmid-mediated AmpC beta-lactamases in Hubei Province of China.

A pepsin modified poly (glycidyl methacrylate-ethyleneglycol dimethacrylate)(poly (GMA-EDMA)) capillary monolith (32 cm ×75 μm,22cm effective lenth)was applied in exploring the interaction between nefo-pam enantiomers and bovine serum albumin (BSA),mode of frontal analysis was selected to measure the binding constant,number of binding sites and the location of binding sites of BSA to both nefopam enantiomers.The opti-mal CEC conditions obtained were a running buffer consisted of 15 mmol /L ammonium acetate at pH 5.5,separa-tion voltage 5.0 kV,detection wavelength 215 nm,injection 10 kV ×6 s,solvent of samples consisted of 50 mmol/L ammonium acetate at pH 7.4.The results indicated that the monolith could provide a satisfactory resolution between the two enantiomers plateaus,BSA in the binding system didn′t disturb the separation or determination of nefopam enantiomers in electrochromatography.The frontal analysis demonstrated that BSA has only one binding site with both enantiomers,the binding constants (K)were 443 L/mol and 527 L/mol,respectively,and the dis-placement experiments indicated that binding site of both isomers to BSA molecule was the Sudlow siteⅡ.

Cluster Analysis ; Fuzzy Logic ; Humans ; Mathematics ; Medicine, Chinese Traditional

Adenocarcinoma, Papillary ; diagnosis ; metabolism ; Carcinoma, Pancreatic Ductal ; diagnosis ; metabolism ; Cystadenoma, Mucinous ; diagnosis ; metabolism ; Gene Expression Regulation, Neoplastic ; Humans ; Mucins ; classification ; genetics ; metabolism ; Pancreas ; metabolism ; Pancreatic Neoplasms ; diagnosis ; metabolism

Objective To investigate the effects of protein kinase C (PKC) on proliferation, extracelluar matrix synthesis and transcriptional factor SOX9 (SRY-related high mobility group-box gene 9) expression of rat growth plate chondrocytes in vitro. Methods Rat costochondral growth plate chondrocytes (RGC) were isolated and cultured. The 1st serum free cultured passage RGCs were treated with 1, 10 and 100 nmol/L phorbol 12,13-dibutyrate (PDBu), PKC agonist, cell morphology were observed with inverted microscopy, cell proliferation, COLLAGEN and GAG synthesis were detected by isotope incorporation COLLAGEN TYPEⅡ and AGGRECAN mRNA transcription and SOX9 expression were revealed by RT-PCR and Western blot.Results 100 nmol/L PDBu treatment made the cell morphology of serum free cultured RGC closed to serum group and inhibited proliferation but promoted COLLAGEN and AGGRECAN synthesis, 3H-TdR、3H-proline and 35S-sulfate incorporation of 100nmol/L PDBu group were as 83%, 52.6% and 146.5% as those of serum free control(P

OBJECTIVE:To observe the efficacy and safety of paroxetine combined with buspirone in the treatment of geriatric depression with anxiety. METHODS:78 patients with geriatric depression with anxiety were randomly divided into control group (39 cases) and observation group (39 cases). Control group received 20-40 mg Paroxetine hydrochloride tablet,within half an hour after breakfast. Observation group additionally received 5-10 mg Buspirone hydrochloride tablet for 3 times. The treatment course for both groups was 8 weeks. Clinical efficacy,HAMD,HAMA and TESS before and after 2,4 and 8 weeks of treatment in 2 groups were observed. RESULTS:The total effective rate in observation group was significantly higher than control group,the difference was statistically significant(P<0.05). HAMD after 4 and 8 weeks,HAMA after 8 weeks in control group,HAMD and HAMA after 2,4 and 8 weeks in observation group were significantly lower than before,and it gradually decreased by time pro-longed,observation group was lower than control group,the differences were statistically significant (P<0.05 or P<0.01). And there was no significant difference in the TESS rating in 2 groups before and after treatment(P>0.05). CONCLUSIONS:The effi-cacy of buspirone combined with paroxetine is superior to paroxetine alone in the treatment of geriatric depression with anxiety, with better safety.

Objective To analysis the effect of compound betamethasone injection combined with radiation on keloid in frontal facial region. Methods 60 cases of keloid patients in zhejiangprovincial people's hospital from January 2014 to January 2016 were selected,all patients were treated with surgery,the patients were randomly divided into observation group and control group,30 cases in each group,radiotherapy was used in the control group,the observation group was treated with local injection of compound betamethasone on the basis of the control group,the therapeutic effects, pain scores, adverse reactions and complications were compared between the two groups. Results The recurrence rate of the observation group was 3.33% which was significantly lower than that of the control group 10%,the improvement rate of the observation group 93.33% was significantly higher than the control group 80%,the difference was statistically significant (P<0.05).one,two and three months after operation,the pain scores in the observation group were significantly lower than those in the control group at the same time point,the difference was statistically significant (P<0.05).The adverse reaction rate was 6.66% (2/30) in the observation group and 23.33% (7/30) in the control group, there was no significant difference between the two groups.The incidences of skin sag, pigmentation, incision delayed union and telangiectasia in the observation group were significantly lower than those in the control group (P<0.05). Conclusion Compound betamethasone injection combined with radiation on the frontal facial keloid patients with significant effect,can significantly reduce the recurrence rate,improve the rate of improvement,reduce pain and complications in patients.

To explore the adhesion of peripheral blood lymphocytes and eosinophils and lung microvascular endothelial cells (LMEC) in asthma. Guinea pigs were divided into asthma model group and control group; the adhesion rates of isolated peripheral lymphocytes, eosinophils and the cultured LMEC monolayers were assayed.The mRNA expressions of VCAM 1 and eotaxin in LMEC were determined by RT PCR.The DNA binding activities of NF ?B and AP 1 of LMEC were detected by electrophoretic mobility shifts assay (EMSA). Under the stimulation with serum of asthma group:①the adhesion rates of lymphocytes, activated eosinophils and LMEC were significantly elevated( P

Objective To investigate the effects of laser irradiation on intracellular ROS(reactive oxidant species),intracellular calcium concentration(_i,and cell membrane integrity in the process of live cell imaging with confocal laser scanning microscopy. Methods The effects of a given laser irradiation on ROS,intracellular calcium concentration(_i and cell viability were revealed respectively by stained ECV-304 with H_2DCFDA,Fluo-4AM and calcein-AM/PI,and visualized and analyzed using ultra view LCI(live cell image)confocal microscopy. Results The irradiation of 488nm laser induced fluorescent intensity of DCF to increase abruptly and attain the climax in about 80 seconds,afterwards the fluorescent intensity fell and returned to the baseline.In the 70 minutes of the irradiation,the fluorescent intensity of intracellular Fluo-4 kept a slightly ascending tendency.The fluorescent intensity of calcein decreased 15minutes after the irradiation,and serval cells were PI positively stained.Conclusion 488nm laser irradiation induces intracellular reactive oxidant species(ROS) and calcium concentration to increase,but there is no significant influence on cell membrane integrity.

Aim To investigate effects of genistein (5, 7, 4′-trihydroxyisoflavone) on rat costochondral growth plate chondrocyte (RGC) collagen and Sox9 expression. Methods Primary cultured RGC, effects of genistein on collagen synthesis, col2a1 mRNA transcription and Sox9 protein expression of 1st passage RGC were detected by isotope incorporation, RT-PCR and western blotting. Results Genistein inhibited collagen synthesis of 1st passage RGC (P