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

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.

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.

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

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.

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

Objective:To develop an HPLC-DAD method for the simultaneous determination of seven alkaloid constitutents, so-phoranholN-oxide, oxymatrine, sophoridine, oxysophocarpine, sophoranol, matrine and sophocarpine in Sophora flavescens Ait. Meth-ods:The chromatographic separation was performed on an Agilent Zorbax C18 column (150 mm × 4. 6 mm, 5μm) with 10 mmol·L-1 NH4Ac(0.1% ammonia, pH=9.0)(A)-acetonitrile-methanol(1 ∶22) containing 20 mmol·L-1NH4Ac(B) as the mobile phase with gradient elution at the flow rate of 1 ml·min-1 . The detection wavelength was set at 220 nm,and the column temperature was maintained at 30 ℃. Results: SophoranholN-oxide, oxymatrine, sophoridine, oxysoph-ocarpine, sophoranol, matrine and sopho-carpine was linear within the range of 0.093-1.860 μg(r =0.999 6) , 0.530-10.600 μg (r =0.999 7), 0.062-1.240 μg (r =0. 999 8), 0. 281-5. 620 μg (r=0. 999 9), 0. 026-0. 520 μg (r=0. 999 8) ,0. 036-0. 720 μg (r=0. 999 7) and 0. 032-0. 640 μg (r=0. 999 6), respectively. The average recovery was 97. 5%, 98. 2%, 99. 0%, 99. 4%, 99. 2%, 98. 2% and 98. 7%, and RSD was 1. 18%, 0. 92%, 1. 43%, 1. 04%, 0. 81%, 0. 43% and 0. 88%(n =6), respectively. Conclusion:The method is convenient, accurate and reproducible in the quality control of multicomponents in Sophora flavescens Ait.

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.