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 explore the effect of failure mode and effect analysis (FMEA) on reducing the unplanned extubation rates for patients with severe craniocerebral injury. Methods FMEA model was used to analyze the main reasons of unplanned extubation patients with severe craniocerebral injury, 7 failure modes with higher RPN scores was selected. The nursing intervention for the fixation of the catheter were improved. The control group with 181 cases of patients without FMEA application were given routine care. FMEA group owning 196 cases were given the optimization process. The incidence rate of unplanned extubation of two groups was compared. Results After improvement of the nursing process of patients with severe craniocerebral injury for fixing the catheter, the RPN of 7 failure modes was lowered. Compared with the control group, the total rate of unplanned extubation in FMEA group decreased to 1.1% (9/832), less than 5.1% (40/779) of the control group. The statistical difference was significant (χ2=22.410, P < 0.01) . The rate of UEX in gastric tube, urinary catheter and central venous catheter were 3.1%(6/196), 1.0%(2/196), 0, lower than 8.8%(16/181), 4.9%(9/181), 3.9%(7/181) in the control group. The statistical difference was significant(χ2=5.718, 5.224, 7.724, P < 0.05). No statistical difference in tracheal cannula and drainage tube of head between the two groups was found(P>0.05). Conclusions The FMEA model can be effective in reducing the rate of unplanned extubation .

The effects of imperatorin (Imp) and iso-imperatorin(Isi) on tumor necrosis factor (TNF) release from mouse peritoneal macrophages were investigated. It was found that Imp and lsi significantly inhibited TNF release from mouse peritoneal macrophages. At the concentration of 10-6～10-4 mol?L-1, the inhibitory effects were presented by Imp and Isi in a dose dependent manner. At 10-4 mol/L TNF release was entirely inhibited by each drug.

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

AIM: To establish the methods for rat costochondral growth plate chondrocyte (RGC) separation and culture and investigate their biological features, thereby provide experimental bases for studying the regulation of chondrocyte proliferation and differentiation. METHODS: RGC were obtained by microdissection and digestion, and cultured in monolayer. Morphological changes of the serial passage of RGC and the cell growth kinectics were observed. The cellular GAG and collagen type Ⅱ expression were detected by histochemistry and ICC. RESULTS: There were more than 98% viable cells in the obtained RGC. The morphology of primary cultured RGC was round or polygon. In this experiment, the sixth passage RGC was still maintained and showed polygonal morphology. The index of duplicatings/day increased in the preceding fourth passage RGC and decreased afterwards. There were more than 95% cells expressed collagen type Ⅱ and alcine blue stained positively in the primary RGC, as the passage number increased, the ratio of collagen type Ⅱ expression and alcine blue positive stained RGC dropped abruptly. CONCLUSION: The separation and culture methods adopted in this study can obtain high pure and viable RGC. The preceding three passage RGC maintains their in vivo phenotype, they are idle experimental materials for studying the regulation of proliferation and differentiation of chondrocyte and tissue engineering.

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.