OBJECTIVE To investigate the produce of extended-spectrum ?-lactamases(ESBLs) and the presence of genotype of the ?-lactamases-encoding genes in Klebsiella pneumoniae isolated from the 98th Hospital of PLA,Huzhou,Zhejiang Province,China.METHODS Twenty-five strains of K.pneumoniae were isolated from the inpatients between Sep 2005 and Apr 2006.ESBLs were tested by phenotypic confirmatory tests recommended by CLSI.Twenty-one kinds of ?-lactamases genes of blaTEM,blaSHV,blaLEN,blaOKP,blaCTX-M-1 group,blaCTX-M-2 group,blaCTX-M-9 group,blaOXA-1 group,blaOXA-2 group,blaOXA-10 group,blCARB,blaPER,blaVEB,blaGES,blaLAP,blaDHA,blaACT/MIR,blaCMY/MOX,blaFOX,blaCMY/LAT,and blaACC were analyzed by PCR and verified by DNA sequencing.RESULTS In 25 strains of K.pneumoniae,the positive,negative,and "uncertainty" rates of ESBLs were 56.0%,20.0%,and 24.0%,respectively.The positive rate of genes of blaTEM,blaSHV,blaCTX-M-1 group,blaOXA-10 group,blaLAP,and blaDHA were 80.0%,4.0%,4.0%,80.0%,4.0% and 32.0%,respectively.The 15 kinds of rest genes were all tested negative.The total positive rate of 21 kinds of ?-lactamases gene was 92.0%.Among them,the blaLAP-2 gene sequence of the HZ12593 strain has been registered in GenBank(GenBank Accession Number: EU529981).CONCLUSIONS There are higher rate of ESBLs-producing strains in K.pneumoniae isolated from the inpatients,and at least 6 kinds of ?-lactamases gene existed.Both genes of blaTEM and blaOXA-10 group are the most common genotypes.Carring blaDHA Gene may influence the result of phenotypic confirmatory test for ESBLs in K.pneumoniae.

Objective To explore the effects of hydrogen sulfide on pulmonary vascular remodeling and its inhibitors in rats with pulmonary hypertension ( PH). Methods Thirty male SD rats were randomly divided into control group ( 10 rats) , model group ( 10 rats) and H2S intervention group ( 10 rats) , PH model was induced by Lilium Wilfordii in model group, on the basis of model group, rats in H2S intervention group were injected with NaHS (56 u,mol/kg) intraperitoneally, while rats in control group were injected with normal saline at the same dose. Four weeks later, the hemodynamic parameters were measured, the right ventricular hypertrophy index (RVHI) was calculated, the pathological changes of pulmonary vessels were detected by HE staining, and the expressions of p38 and c-Jun N-terminal kinase( JNK) proteins in the mitogen-activated protein kinase (MAPK) family were detected by Western blotting and Real-time PCR. Results There were significant differences in hemodynamics, RVHI, wall thickness as a percentage of vessel diameter ( WT) % , pulmonary vessel wall area as a percentage of vascular cross-sectional area( WA) % , p38 and JNK in each group (P<0. 05). The expression levels of MSAP, MPAP, RVHI, WT%, WA%, p38 mRNA and JNK mRNA in the model group and H2S intervention group were significantly higher than those in the control group (P<0. 05) , while the levels of MSAP , MPAP , RVHI, WT% , WA% , p38 mRNA and JNK mRNA in H2S intervention group were significantly lower than those in the model group (P<0. 05). The pulmonary artery morphology showed that the wall thickness and lumen stenosis of the model group and the H2S intervention group increased compared with the control group, but the lumen thickness and lumen stenosis of the H2S intervention group were significantly reduced compared with the model group; Western blotting showed that the expressions of p38 and JNK in model group and H2S intervention group were higher than those in control group, while the expressions of p38 and JNK in H2S intervention group were lower than those in model group. Conclusion H2S can improve hemorheology, right ventricular hypertrophy index, alleviate pulmonary artery wall thickening and lumen stenosis, and inhibit pulmonary vascular remodeling in PH rats. Its mechanism may be related to the down-regulation of JNK and p38 protein expression in MAPK signaling pathway by H2S.

OBJECTIVETo explore the outcomes after surgical treatment of esophagogastric junction carcinoma (EGJC).

METHODSOne hundred and eighty-five patients with EGJC undergoing surgery from October 2000 to September 2006 at the Cancer Hospital of Shantou University were reviewed retrospectively. The clinical outcomes were compared between transthoracic and transabdominal approach.

RESULTSOf the 185 patients, 133 underwent operation via transthoracic approach and 52 via transabdominal approach. The postoperative complication rates were 10.5%(14/133) and 11.5%(6/52) and the 1-, 3-, 5-year overall survival rates were 83.9%, 44.5%, 32.9% and 86.0%, 38.0%, 30.0% in transthoracic and transabdominal groups respectively, and the difference were not statistically significant (both P>0.05).

CONCLUSIONSurgical approach should be individualized for EGCJ.

Adenocarcinoma ; surgery ; Aged ; Esophagogastric Junction ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo observe effect and mechanism of n-Butanol lysate of alcohol extracts from Actinidia rufa root (monomer of R6,R8).

METHODTunel, Wright's stain with Giemsa's stain dyeing, and Hoechst 33258-PI double dyeing assay were used to detect the apoptosis of SGC7901 tumor cells treated with R6, R8. The SGC7901 tumor cells were randomly divided into control group and two treatment groups administered 0.05 g x L(-1) R6, R8, respectively, for 72 h). FCM assay was used to detect the apoptosis. Agarose electrophoresis assay was used to detect DNA strand break of tumor cells and reveal anti-tumor action mechanism.

RESULTThe apoptosis percentage of the tumor cell in 24 h, 48 h, 72 h was (17.08 +/- 2.78)% , (29.68 +/- 2.96)%, (52.46 +/- 3.81)%; (14.75 +/- 2.14)%, (27.35 +/- 3.79)%, (45.64 +/- 5.24)%, respectively, for the treatment group, significantly higher than that in the control group (1.94 +/- 1.55)%, (2.78 +/- 1.84)%, (11.8 +/- 2.79)% (P < 0.01) by tunnel assay. Wright's stain with Giemsa's stain dyeing assay, Hoechst 33258-PI and FCM double dyeing assay showed same action. R6 and R8 had the effect of inducing the DNA histogram of tumor cells (P < 0.01).

CONCLUSIONThe anti-tumor mechanisms may be associated with inducing the injury of DNA and stimulating apoptosis.

Actinidia ; chemistry ; Apoptosis ; drug effects ; Cell Line, Tumor ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Flow Cytometry ; Humans ; Immunohistochemistry ; Plant Roots ; chemistry

Objective To investigate the status of genotype of the KPC(Klebsiella pneumoniae carbapenemase)-encoding genes in Pan-resistant K. Pneumoniae, isolated from the 98th Hospital of People' s Liberation Army, Huzhou district, Zhejiang province, China. Methods 19 strains of Pan-resistant K. Pneumoniae were isolated from the inpatients between November, 2008 and July,2009. Phenotypic confirmatory test for suspected carbapenemases production were carried out by Modified Hodge test. Carbapenemase gene of blaKPC was analyzed by PCR and verified by DNA sequencing. Results In 19 strains of K. Pneumoniae, the positive rates of Modified Hodge test and gene of blaKPC were both 100.0%. These genes all belonged to blaKPC-2 subtype confirmed by nucleotide sequence analysis. Among them, the blaKPC-2 gene sequence of the HZ001 strain (its original serial number was HZ9871 ) had been registered in GenBank (GenBank Accession Number: GU086225).Conclusion All of the Pan-resistant K. Pneumoniae isolated from the inpatients harbored blaKPC-2 type carbapenemases gene and causing an outbreak in a hospital. Carbapenemases that producing type KPC-2 might be the major reason which causing the resistance to Carbapenems antibiotics.

Objective: : To study the physiochemical characteristics of podophyllotoxin (PPT) conjugated stearic acid grafted chitosan oligosaccharide micelle (PPT-CSO-SA), and evaluate the ability of the potential antineoplastic effects against glioma cells. Methods: : PPT-CSO-SA was prepared by a dialysis method. The quality of PPT-CSO-SA including micellar size, zeta potential, drug encapsulation efficiency and drug release profiles was evaluated. Glioma cells were cultured and treated with PPT and PPT-CSO-SA. The ability of glioma cells to uptake PPT-CSO-SA was observed. The proliferation of glioma cells was determined by 3-[4, 5-dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay. The apoptosis and morphology of U251 cells were observed by 4’,6-Diamidino-2-phenylindole dihydrochloride (DAPI) dye staining. Cell cycle analysis was performed by flow cytometry. The migration ability of U251 cells was determined by wound healing test. Results: : PPT-CSO-SA had nano-level particle size and sustained release property. The encapsulation efficiency of drug reached a high level. The cellular uptake percentage of PPT in glioma cells was lower than that of PPT-CSO-SA (p<0.05). The inhibitory effect of PPT-CSO-SA on glioma cells proliferation was significantly stronger than that of PPT (p<0.05). The morphologic change of apoptosis cell such as shrinkage, karyorrhexis and karyopyknosis were observed. The percentage of U251 cells in G2/M phase increased significantly in the PPT-CSO-SA group compared with PPT group (p<0.05). Compared with the PPT group, the cell migration ability of the PPT-CSO-SA group was significantly inhibited after 12 and 24 hours (p<0.05). Conclusion : PPT-CSO-SA can effectively enhance the glioma cellular uptake of drugs, inhibit glioma cells proliferation and migration, induce G2/M phase arrest of them, and promote their apoptosis. It may be a promising anti-glioma nano-drug.

Antigens, CD ; genetics ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Tetraspanin 24

OBJECTIVETo investigate the influence of insulin growth factor-I (IGF-I) on apoptosis of cardiomyocytes subjected to ischemia and hypoxia and its possible mechanism.

METHODSCardiomyocytes were cultured in vitro, and randomized into hypoxia group, treatment group (T, the cells were treated with IGF-1 before subjected to hypoxia and ischemia) and control group (C, normal cardiomyocytes as controls). Changes in the OD value of cell apoptosis, mitochondrial membrane potential and relative amount of phospho-Akt protein were observed at different time-points by ELISA, laser scanning with TMRE staining and Western blot, respectively.

RESULTSThe OD value of cell apoptosis in control group was 0.18 +/- 0.03, while that in hypoxia group was gradually increased to 0.33 +/- 0.05, 0.61 +/- 0.06, 1.17 +/- 0.08, 2.25 +/- 0.11, respectively at 1, 3, 6, 12 post-hypoxia hours (PHH), showing an increasing tendency (P < 0.01). The OD values of cell apoptosis in T group were 0.26 +/- 0.04, 0.49 +/- 0.05, 0.84 +/- 0.06, 1.63 +/- 0.09, respectively, which were obviously lower than those in hypoxia group (P < 0.05 or P < 0.01). The mitochondrial membrane potential (Dymt) values in hypoxia group at 6 and 12 PHH were 18.7 +/- 5.1 and 6.3 +/- 1.9, respectively, which were obviously lower than that in control group (40.2 +/- 10.1, P < 0.01). The DYmt in T group at 6 and 12 PHH were 28.8 +/- 6.2, 12.5 +/- 3.1, respectively, which were obviously higher compared with those in hypoxia group (P < 0.05). The amount of phospho-Akt protein was increased by IGF-I administration.

CONCLUSIONIGF-I exhibits an anti-apoptotic effect on cardiomyocytes subjected to ischemia and hypoxia, and this may be related to the activation of PI3K/Akt signal pathway and stabilization of mitochondrial membrane potential.

Animals ; Apoptosis ; Cell Hypoxia ; Cells, Cultured ; Hypoxia ; metabolism ; Insulin-Like Growth Factor I ; pharmacology ; Ischemia ; metabolism ; Membrane Potential, Mitochondrial ; Myocytes, Cardiac ; cytology ; Proto-Oncogene Proteins c-akt ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

[Objective]To investigate the effect of KIF23 gene expression on the proliferation,migration and invasion of human hepatocellular carcinoma HepG2 cells in vitro,and to explore the possible mechanism.[Methods]The KIF23 siRNA was transfected into HepG2 cells by lipofectamine 3000.The expression of KIF23 mRNA and protein in HepG2 cells was de-tected by qRT-PCR and Western blot.The effect of silencing KIF23 on the proliferation of HepG2 cells was studied by CCK-8 assay and plate clone formation assay.The tumor cell abilities of migration and invasion after transfection were measured by scratch assay and Transwell assay.The expression of protein kinase B(PKB/Akt)and phosphorylated Akt(p-Akt)protein in HepG2 cells transfected with KIF23-siRNA2 was detected by Western blot.[Results]KIF23-siRNA could effectively si-lence the expression of KIF23 mRNA and protein in HepG2 cells(P<0.01).The results of CCK-8 assay,plate clone forma-tion assay,scratch assay and Transwell assay demonstrated that the cell proliferation,migration and invasion ability of the KIF23-siRNA2 interference group were significantly inhibited,compared to the negative control group and the blank control group(P<0.05).The expression level of total Akt protein in HepG2 cells was not changed,but the expression level of phos-phorylated Akt protein was down-regulated(P<0.05).[Conclusions]KIF23 may promote the proliferation,migration and in-vasion of human hepatocellular carcinoma cells by activating Akt signal transduction pathway.KIF23 is expected to be a new target for gene therapy of hepatocellular carcinoma.

Genes encoding nucleocaspid (N) and membrane (M) protein of SARS coronavirus were obtained by RT-PCR and were cloned into expression vector pET22b and pBV222. DNA sequencing showed that the genes cloned from a patient in Beijing were identical to the gene sequences from reported Toronto strain. The genes were over-expressed in E. coli either as inclusion body or as soluble form. The recombinant proteins were purified by ion-exchange, or ion-exchange followed by metal chelate affinity chromatography. The recombinant N protein was demonstrated highly antigenic and could be employed as antigen to detect SARS antibodies in ELISA system for SARS diagnosis.
Chromatography, Affinity ; Chromatography, Ion Exchange ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; metabolism ; Nucleocapsid Proteins ; genetics ; isolation & purification ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus ; genetics ; metabolism ; Viral Structural Proteins ; genetics ; isolation & purification ; metabolism