Objective To investigate the resistance mechanisms and epidemiology of Enterobacteriaceae isolated from clinical with reduced susceptibility to imipenem or meropenem.Methods 18 strains of Enterobacteriaceae with reduced carbapenem susceptibility were collected during January to August in 2010.The MICs of these strains were determined using automated microbial identification system.ESBLs,AmpC and KPC were tested using the agar dilution method.PCR amplification and DNA sequence were performed to analyze the KPC genes,PFGE was used to examine the molecular epidemiology.Results All 18 strains were detected ESBLs and AmpC,14 strains were detected KPC-2.3 strains with EDTA paper method positive may produce other metal carbapenem,in which 2 strains harbor KPC-2.PFGE types indicate that there were six genotypes among 15 strains of Klebsiella pneumoniae.Conclusion Plasmid-mediated KPC-2 was the main reason which makes Enterobacteriaceae reducing carbapenem susceptibility and causes short-term epidemic in hospital.Clinical strains harboring KPC-2 gene may carry multiple resistance genes meanwhile.

Background Oxidative damage may cause the functional dysfunction and death of retinal vascular endothelial cells(RVECs),and further leads to the development of retinal vascular diseases.Fufang xueshuantong has a therapeutic effect on retinal vascular diseases,but little is known about its molecular mechanism.Objective The goal of this study was to investigate the protective effects and mechanism of fufang xueshuantong on injury of human RVECs induced by tert-butyl hydroperoxide(t-BHP).Methods Human RVECs were isolated from healthy donor eyes and primarily cultured and then identified by flow cytometry.The third to fifth generations of cells were used in this experiments.The fufang xueshuantong solution of 0.0625,0.1250,0.2500,0.5000 and 1.0000 g/L were added in the cuhure plate with 5 × 104/L cells respectively in the experimental groups,and t-BHP of 75,100,200 and 300 μ.mol/L were added in the model control groups.MTT was used to detect the A490and survival rate of RVECs.The apoptotic rate and death rate of the cells were evaluated by double staining of Annexin V-FITC/PI.Morphology of human RVECs were examined using invert microscopy and Hoechst33258 staining.The expressions of nitro tyrosine (a marker of oxidative damage of protein)and 8-OHdG(a marker of oxidative damage of DNA)in human RVECs were assessed by the immunofluorescence staining.Western blot was used to detect the expressions of nuclear factorkappa B(NF-KB),p53,bcl-2 and bax after 6,12,24 hours t-BHP action.This study was approved by the Ethic Committee of Zhongshan Ophthalmic Center.Results No significant difference was found in A490value among the normal control group,0.0625,0.1250,0.2500,0.5000 and 1.0000 g/L fufang xueshuantong groups(F =1.989,P＞0.05).The survival rates of the cells were lower in 75,100,200 and 300 μmol/L t-BHP groups compared with corresponding fufang xueshuantong groups(t =14.57,13.82,21.51,32.64,P＜ 0.01).The percentages of normal cells were evidently lower in 75,100,200 and 300 μmol/L t-BHP compared with corresponding fufang xueshuantong groups(t=14.908,5.495,17.165,26.330,P＜0.01).The numbers of deformation and death of the human RVECs increased as the elevated concentration of t-BHP,but those in fufang xueshuantong groups were less than the t-BHP groups under the invert microscopy.Compared with t-BHP groups,the expressions of nitro tyrosine,8-OHdG,NF-KB,p53 and bax were lower but the expression of bcl-2 was higher in human RVECs with the statistically significant differences(P＜0.05).Fufang xueshuantong at the concentration of 0.2500 g/L showed maximally protective effect on human RVECs.Conclusions Fufang xueshuantong protects human RVECs against the t-BHP-induced injury through downregulating the expression of NF-kB,p53,bax and up-regulating the express of the bcl-2 protein.

A novel serine protease with high purity was extracted from the venom of Agkistrodon hlays Pallas using monoclonal antibody affinity chromatography. This protease releases bradykinin and has arginine esterase activity without being activated. After purification, its hydrolytic activity exceeded 800 U/mg, far higher than its counterparts from mammalian sources. The purity of the kininogenase could exceed 95%. The acute toxicity and the long-term toxicity of this kallikrein was studied for its potential clinical application. The maximum tolerance dose in adult was 150,000 times greater than the maximum applied dose, and long-term administration at the dose 50 times of allowed clinical dose did not obviously after the animals' body weight, survival condition, liver function, renal function, or blood routines, suggesting the extremely low toxicity of the kallidrein.
Agkistrodon ; Animals ; Bradykinin ; metabolism ; Crotalid Venoms ; toxicity ; Kallikreins ; toxicity ; Maximum Tolerated Dose ; Mice ; Mice, Inbred BALB C ; Serine Endopeptidases ; toxicity ; Toxicity Tests, Acute

OBJECTIVEConditioned taste preference (CTP) is a taste learning reflex by which an animal learns to prefer a substance which tastes not well and has been studied with much interest in recent years. However, the neural substrates of CTP are less known. This study aimed to determine the possible neural path- ways of CTP and whether serum leptin level and the leptin receptor (OB-Rb) in the hind brain are involved following CTP formation.

METHODSWe established CTP of quinine in rats with a 2-bottle preference test. The serum leptin concentrations were detected, the expression of c-fos in the rat brain was tested to determine the nuclei in relation with establishment of CTR Finally, the OB-Rb mRNA expression was examined by RT-qPCR assay in parabrachial nucleus (PBN) and the nucleus of the solitary tract (NST) of the hind brain.

RESULTSCompared with control group, the level of serum leptin was higher in the CTP group (4.58 ± 0.52 vs 1.67 ± 0.25 µg/L, P < 0.01); increased c-fos positive cells were found in the anterior hypothalamus (AH, 221.75 ± 4.96 vs. 178.50 ± 6.63 cells/mm², P < 0.05), the basal lateral amygdala (BLA, 70.75 ± 6.17 vs 56.50 ± 3.62 cells/ mm², P < 0.05) and the nucleus of the solitary tract (NST, 41.25 ± 1.32 vs 32.50 ± 1.02 cells/mm², P < 0.05). But in ventromedial nucleus of the hypothalamus (VMH, 20.75 ± 2.73 vs 38.5 ± 1.54 per 1 mm², P < 005), PBN (21.50 ± 2.24 vs 36.25 ± 1.49 cells/mm², P < 0.05) and the central nucleus of the amygdala (CeA, 22.25 ± 1.53 vs 35.50 ± 2.11 cells/mm², P < 0.05), the number of c-fos positive cells was decreased in the CTP group. In addition, we found OB-Rb mRNA expression in PBN of CTP group rats was higher than that of control group (0.95 ± 0.055 vs 0.57 ± 0.034, P < 0.05), while there was no significant difference of OB-Rb mRNA expression in NST between the two groups.

CONCLUSIONNuclei AH, BLA, NST, VMH, PBN and CeA participate in the formation of CTP. Leptin and its receptor in PBN may be involved in the formation and maintenance of CTP.

Animals ; Conditioning (Psychology) ; Leptin ; blood ; Rats ; Receptors, Leptin ; physiology ; Rhombencephalon ; physiology ; Taste ; physiology

Objective To identify the expression of sphingosine-1-phosphate receptor (S1PR) subtypes, C-myc and His tag proteins of human umbilical vein endothelial fusion cell line, EA.hy926 and human umbilical vein endothelial cells (HUVEC), CRL-1730 for studying the function of apolipoprotein M (ApoM)-S1P axis. Methods Two kinds of cells (EA. hy926 and CRL-1730) were cultured to reach the density of 60%-70% in vitro. Immunofluorescence technique was em?ployed to investigate the expressions of coagulation factorⅧ(FⅧ), ApoM, S1PR1-S1PR5, C-myc and His tag proteins. Re?sults (1) Two kinds of cells both expressed FⅧand ApoM. FⅧpresented scattered particle distribution in CRL-1730, while uniform distribution in EA.hy926. However, ApoM was strongly expressed and widely distributed in cytoplasm of two kinds of cells. (2) S1PR1-3 can be detected on their membrane other than S1PR4 and S1PR5. S1PR1 was highly expressed but S1PR2 and S1PR3 were in a low level expression. (3) Two kinds of cells both expressed C-myc and His tag proteins in cytoplasm. Conclusion Two kinds of cells have the properties of endothelial cells and can express FⅧ, ApoM, C-myc and His tag proteins. It is not suitable for choosing C-myc and/or His tag–conjugated recombinant ApoM to study the fuction of ApoM-S1P axis with these two kinds of cells.

AIM: To investigate the relationship among MCP-1 and monocyte chemoattract protein activity (MCA) and pathogenesis of lung cancer. METHODS: 173 patients were involved in the study and divided into three groups: group A: lung cancer group (60 patients); group B: benign lung disease group (55 patients) and group C: healthy control group (58 patients). MCP-1 level and MCA in bronchoalveolar lavage fluid (BALF) were measured. RESULTS: The concentration of MCP-1 and MCA in BALF in group A were much higher than those in group B and group C (P

OBJECTIVETo investigate the effect of bufalin on nucleus-mitochondria localization of human telomerase reverse transcriptase(hTERT) by exploring its effect on proliferation and apoptosis in human esophageal squamous carcinoma EC9706 cells.

METHODSEC9706 cells were treated with bufalin at various concentrations, and then the cell growth inhibition of EC9706 cells was examined by CCK-8 assay and the 50% inhibitory concentration (IC(50)) was calculated.Cell cycle analysis was performed by flow cytometry with PI staining, and nucleus morphology of apoptosis were observed by fluorescence microscopy with Hoechst 33342 staining. The apoptotic index was measured by flow cytometry with Annexin V-FITC/PI double staining. hTERT subcellular localization and protein expression were determined by Western blotting and multiple immunofluorescence labling combined with laser confocal scanning microscopy.

RESULTSThe proliferation of EC 9706 cells was significantly inhibited by bufalin along with the increase of processing time and concentrations (p<0.01). After the EC9706 cells were exposed to 100 nmol/L bufalin,the number of cells gradually decreased in G(1) phase and increased in S and G(2)/M phases(p<0.05). The typical nucleus morphological changes of apoptosis were observed and the apoptotic index was increased(p<0.01). The expression of hTERT decreased in nucleus but increased in mitochondria(p<0.05).

CONCLUSIONSBufalin can inhibit the proliferation of human esophageal squamous carcinoma EC9706 cells in a time- and dose-dependent manner. It can arrest cell cycle in S and G(2)/M phases and induce the apoptosis of EC 9706 cells. hTERT is localized in both nucleus and mitochondria,and can be partially translocated from nucleus to mitochondria during the bufalin-induced apoptosis.

Apoptosis ; drug effects ; Bufanolides ; pharmacology ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Esophageal Neoplasms ; metabolism ; pathology ; Humans ; Telomerase ; metabolism

The clinical application of Chinese patent medicines has suffered sever problems and required guidelines for clinical practices. Currently, the expert consensus method is more suitable for formulating clinical practice guidelines of Chinese patent medicines than the evidence-based method. However, there remain problems in the application of the expert consensus method. This study proposed a derivative expert consensus method--a method for formulating clinical practice guidelines of common Chinese patent medicines based on clinical practices, and introduced the method in terms of research thought, methodology and implementation procedure.
Evidence-Based Medicine ; standards ; Humans ; Nonprescription Drugs ; standards

In order to develop a rapid detection kit for novel avian influenza virus (AIV) subtype H7N9, two sets of specific primers and probes were designed based on the nucleotide sequences of hemagglutinin antigen (HA) and neuraminidase antigen (NA) of novel H7N9 virus (2013) available in GenBank to establish the method of TaqMan probe-based multiplex real-time RT-PCR for rapid detection of AIV subtype H7N9. The primer and probe of HA were for all H7 subtype AIVs, while the primer and probe of NA were only for novel N9 subtype AIVs. The results showed that this method had high sensitivity and specificity. This method was applicable to the testing of positive standard sample with a minimum concentration of 10 copies/microL; it not only distinguished H7 subtype from H1, H3, H5, H6, and H9 subtypes, but also distinguished novel N9 subtype from traditional N9 subtype. A total of 2700 samples from Zhuhai, China were tested by this method, and the results were as expected. For the advantages of sensitivity and specificity, the method holds promise for wide application.
Animals ; Birds ; virology ; Influenza A Virus, H7N9 Subtype ; genetics ; isolation & purification ; physiology ; Influenza in Birds ; prevention & control ; virology ; Real-Time Polymerase Chain Reaction ; methods ; Species Specificity ; Taq Polymerase ; metabolism ; Time Factors

OBJECTIVETo evaluate the effect of irbesartan on the proliferation, apoptosis, and VEGF mRNA expression of human umbilical vein cell line EA.hy926 in vitro.

METHODSThe human umbilical vein cell line EA.hyY926 were treated with various concentrations of irbesartan for 24 h. The cell proliferation after the treatment was detected by CCK8 assay, flow cytometry and FITC Annexin V/PI kit were used to detect changes in the cell apoptosis. RT-PCR was used to evaluate the expression of VEGF mRNA.

RESULTSThere were no changes in cell shape with various concentration of irbesartan. CCK-8 assay showed a greater rate of the cell proliferation in irbesartan group than that in control group with a dose-independent manner after 24 h treatment. After incubation with irbesartan, cell proliferation rate was significant (P < 0.05). FCM analysis showed no significantly changes in the cell apoptosis. Irbesartan increased the proliferation of EA.hy926 cells. At concentration of 1 x 10(-4), 1 x 10(-5), 1 x 10(-6) mol/L, VEGF mRNA expression enhanced either (P < 0.05).

CONCLUSIONIrbesartan could promote the proliferation and up-regulated VEGFmRNA expression in EA.hy926 cell line. This result suggested that in addition to antihypertensive effect, angiotensin receptor antagonist might be a novel therapeutic approach to chronic ischemic heart disease as heart failure.

Apoptosis ; drug effects ; Biphenyl Compounds ; pharmacology ; Cell Line ; Cell Proliferation ; drug effects ; Humans ; RNA, Messenger ; genetics ; Tetrazoles ; pharmacology ; Umbilical Veins ; cytology ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism