The bacterial strain CSS-4-2 isolated from the spleen of Carassius auratus of penze(♀) ? Cyprinus acutidorsalis (♂) with septicemia was proved to b e a pathogen by artificial infection. This bacteria has short rod-shaped cells , Gram negative and motile, with single polar flagellum, and with not capsule. No spore was formed , and it was resistant to 0/129. It could utilize glucose, mal t ose ,mannite, sucrose and inositol, but could not utilize malonate, lactose, xyl ose, raffinose, sorbin and adonitol. Oxidase and arginine dihydrolase was posit ive.While urease, lysine decarboxylase and ornthine decarboxylase was negative .Bacteriological identification by VITEK-AMS-60 showed the strain CSS-4-2 w as Aeromonas caviae. Drug sensitivity assay showed that this strain was sen sitive to s treptomycin, tobramycin, chloromycetin, rifampicin, norfloxacin and nitrofuranto in, but resistant to penicillin G.

The growth property of pathogenic bacterium Aeromonas caviae strain CSS-4-2 which was isolated from the spleen of Carassius auratus of penze(♀) ? Cyprinus acutidorsalis (♂) that diseased septicemia was studied.The growth curve of CSS-4-2:lag phase was in 0～3h,log phase was in 3～30h, stationary phase was in 33～36h,decline phase was after 36h when CSS-4-2 was cultured shaking in tryptone liquid medium. It's optimum growth temperature ranges from 25℃ to 30℃, optimum growth pH ranges from 5 to 10, and optimum salinity ranges from 0% to 1% Nacl concentration .

Tumor genesis and development often result from deregulation of important biological pathways at the gene expression level. Although there has been much work focused on searching gene sets using gene expression data or other prior information, proper statistical testing of the gene sets is still an open question. Most studies have expanded the testing method of a single gene into the gene sets. Parametric statistical analysis of gene sets ( p-SAGE ) was presented for determining the significant gene sets or pathways associated with a phenotype of interest. The method was applied to brain tumor experiments to identify many gene sets. Some of the newly discovered gene sets were related to signal transduction and immunity. This simple and effective method gives useful biologically meaningful results.

A new phenethanol, (2'R)-4-(2', 3'-dihydroxy-3'-methyl-butanoxy)-phenethanol (1), along with other eleven known benzene derivatives (2-12) were isolated from the roots, stems and leaves of Clausena excavata (Rutaceae). Compounds 3 and 4 are new natural products, and compounds 5-8, 10-12 were isolated from C. excavata for the first time. Their structures were elucidated on the basis of MS, 1D and 2D NMR spectroscopic analyses including HSQC, COSY and HMBC experiments. 1 was tested for its cytotoxicities against A549, HeLa and BGC-823 cancer cell lines, and antimicrobial activities against Candida albicans and Staphylococcus aureus. The results showed that 1 did not exhibit cytotoxic and antimicrobial activities.
Benzene Derivatives ; chemistry ; Candida albicans ; drug effects ; Cell Line, Tumor ; Clausena ; chemistry ; HeLa Cells ; Humans ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plant Leaves ; chemistry ; Plant Roots ; chemistry ; Plant Stems ; chemistry ; Staphylococcus aureus ; drug effects

To investigate monoterpenes and sesquiterpenes of the stems and leaves of Clausena excavata, an AcOEt fraction of the methanol extract was subjected on column chromatographies including silica gel and RP-18, as well as preparative HPLC. The structures of compounds isolated were identified on the basis of spectroscopic data as excamonoterpene (1), (6R, 9S)-9, 10-dihydroxy-4-megastigmen-3-one (2), (3R, 6R, 7E) -3-hydroxy-4, 7-megastigmadien-9-one (3), (3S) -3-hydroxy-7, 8-dihydro-beta-ionone (4), (3S, 5R, 6S) -3-hydroxy-5,6-epoxy-beta-ionone (5), (6R, 9R) -9-hydroxy-4-megastigmen-3-one (6), (3S, SR) -dihydroxy-6, 7-megstigmadien-9-one(7), (-)-loliolide(8), caryolane-1, 9alpha-diol(9) and 2, 6-dihydroxyhumula-3 (12), 7 (13), 9(E)-triene (10), were isolated from the stems and leaves of C. excavata. Compound 1 is a new monoterpene, named as excamonoterpene. Compounds 2-10 were isolated from this plant for the first time.
Chromatography, High Pressure Liquid ; methods ; Clausena ; chemistry ; Magnetic Resonance Spectroscopy ; Methanol ; chemistry ; Molecular Structure ; Monoterpenes ; analysis ; chemistry ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Sesquiterpenes ; analysis ; chemistry ; Spectrometry, Mass, Electrospray Ionization

Diagnosis, Differential ; Hemangiopericytoma ; metabolism ; pathology ; Hemangiosarcoma ; metabolism ; pathology ; Histiocytoma, Malignant Fibrous ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Leiomyosarcoma ; metabolism ; pathology ; Nerve Sheath Neoplasms ; metabolism ; pathology ; Neuroectodermal Tumors, Primitive ; metabolism ; pathology ; Rhabdoid Tumor ; metabolism ; pathology ; Rhabdomyosarcoma ; metabolism ; pathology ; Soft Tissue Neoplasms ; metabolism ; pathology ; Urinary Bladder ; metabolism ; pathology

OBJECTIVETo investigate the effects of advanced glycosylation end products (AGEs) modified bovine serum albumin (AGE-BSA) on the expression of reactive oxygen species (ROS) and monocyte chemoattractant protein-1 (MCP-1) in human renal mesangial cells (HRMCs).

METHODSHRMCs were cultured in vitro with medium containing different doses of AGE-BSA or BSA (50,100, 200, 400 mg/L) for 48 hours, or with AGE-BSA (200 mg/L) for different times (12, 24, 48, 72 h). Immunocytochemistry assay was used to estimate the protein level of RAGE. The ROS in cells were measured by flow cytometry and the mRNA expression of MCP-1 were analyzed by semi-quantiative reverse transcription-polymerase chain reaction (RT-PCR) after treatment with AGE-BSA or BSA.

RESULTSThe protein level of RAGE was upregulated in the HRMCs with AGE-BSA. The expression of ROS and MCP-1 significantly enhanced by incubation of AGE-BSA in a time- and dose-dependent manner. The effects of AGE-BSA-induced up-regulation of ROS and MCP-1 level was significantly blocked by neutralizing antibodies to RAGE, while the expression of ROS and MCP-1 stood nearly unchanged after cultured with huamn IgG.

CONCLUSIONThe expression of ROS and MCP-1 in HRMCs is induced by AGE-BSA through RAGE, which may have potential effects in the pathgenic mechanism of diabetic nephropathy.

Cells, Cultured ; Chemokine CCL2 ; metabolism ; Glycation End Products, Advanced ; pharmacology ; Humans ; Mesangial Cells ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Reactive Oxygen Species ; metabolism ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; Serum Albumin, Bovine ; pharmacology

Objective To study the growth inhibition,apoptosis induction effects of cinobufagin(CB)on human osteosarcoma(OS) cell line U2OS,MG63 and SaO2 in vitro and the underlying mechanism of action of cinobufagin in OS cells.Methods Cell viability was assessed by MTT assay.Cell-cycle status,apoptosis-inducing effects were evaluated by flow cytometry,fluorescent staining and DNA fragmentation assays.Inhibitors of apoptosis proteins (IAPs) and Bcl-2 family proteins including Bax,cleaved-PARP,xIAP,cIAP-1,survivin and p65 were tested by Western blot.Results MTT assay showed that CB could inhibited the growth of U2OS,MG63 and SaO2 cells in a dose-and time-dependent manner.The 48 h IC50 of CB on U2OS,MG63 and SaO2 cells were (104.83± 16.96) nmol/L,(47.07±7.5) nmol/L,and (136.72±10.08) nmol/L respectively.The induction of G2/M cell-cycle arrest was seen in the cells treated with CB.After cells were cultured for 12 h in the presence of 100 nmol/L CB,the percentages of cells in the G0/G1 phase were decreased,while G2/M phase were increased in U2OS,MG63 and SaOS2 cells,respectively.The results showed CB inhibited the proliferation of osteosarcoma cells through blocking the cell cycle in G2/M phase.Induction of apoptosis was confirmed by Hoechst 33258 and Annexin V/PI staining.After treating with 100 nmol/L CB for 48 h,the extents of apoptosis were 33.6％±6.4％,36.4％±7.8％ and 29.3％±5.1％,respectively.These results indicate that the anti-tumor activity of cinobufagin in osteosarcoma cells was due to a G2/M cell cycle arrest and apoptosis inducing effect.Western blot showed that CB could induce the apoptosis related family proteins Bax,cleaved-PARP up-regulation,xIAP,cIAP-1,survivin and p65 downregulation in OS cells.Conclusion CB can inhibit the cell viability and induce G2/M cell cycle arrest and apoptosis in U2OS,MG63 and SaO2 cells.The apoptosis-inducing effect of CB is confirmed by the regulation of apoptosis related proteins IAPs and Bcl-2 in vitro.

Objective To observe the effect of leukocytapheresis(LCAP)on the coagulation,fibrinolysis system and lung injury in the endotoxemia dog and explore the mechanism in the endotoxin-induced lung injury dog. Methods Endotoxemia-induced model in dogs was established by administration of lipopolysaccharide(LPS,2 mg/kg).Separation of the leucocytes wag performed with the automated continuous flow blood cell separators.A total of 30 male mongrel dogs were randomly divided into LPS group(group L,only injected with LPS,with no LCAP),sham LCAP group(group S,received sham LCAP at 12-14 hours after administration of LPS)and LCAP treatment group(group T,received LCAP at 12-14 hours after administration of LPS),10 dogs per group.The dynamic changes of the activated protein C(APC),soluble thrombomodulin and plagminogen activator inhibitor-1 in the serum were measured at 0 hour before LPS administration,at 2,6,12,14,16,24 and 36 hours after administration of LPS.Results Through LCAP,there found the following four results:(1) the APC level in the serum of the group T wag(50.805±4.422)μg/ml and(40.480±2.993)μg/ml at 14 hours and 16 hours respectively,which were significantly higher than(45.881±4.024)μml and(35.935±4.057)μg/ml in the group L(P<0.05).(2)The expressions of soluble thrombomadulin in the group T was (9.688±O.914)μml and(10.492±O.865)μg/ml at 14 hours and 16 hours respectively,which was statistically lower than(11.005±0.854)μg/ml and(12.04±0.954)ug/ml in the group L(P<0.05).(3)Thelevel of plagminogen activatorinhibitor-1 in the group T was lower than that in the group the group T Wag statistically lower than that in the group L(ALI/ARDS occurred in 2 and 7 dogs of the groups T and L respectively within 36 hours after infusion of LPS.P<0.05). Conclusions At the decrease the incidence of acute lung injury partly due to its role in improving the function of coagulation and fibrinolysis.

Murine norovirus (MNV) was first discovered in mice in 2003. MNV is a member of the genus Norovirus in the family Caliciviridae. It is one of the most important and prevalent pathogens of laboratory mice, and almost all mouse strains are susceptible to MNV infection. In this study, a MNV strain was isolated from the cecal contents of infected mice and identified by the cytopathic effect (CPE) assay, virus plaque assay, 50% tissue culture infectious dose (TCID50) assay, electron microscopy, indirect immunofluorescence assay (IFA) and nucleotide sequencing. On infection, the RAW264.7 cell line showed obvious cytopathic effects within 24 to 48 hours post-inoculation, as infected cells became rounded, bright and shrunken, with ultimate disintegration of the cell sheet. After the isolation of the MNV virus, the virus was plaque-purified in RAW264.7 cells. The TCID50 of the virus was 10(5.25/0.1 mL. Electron microscopic observations of the purified virus showed the presence of spherical and non-enveloped viral particles that were 30 to 35 nm in diameter. According to the identification results, the isolate was named as MNV Guangzhou/K162/09/CHN. Thereafter, five overlapping gene fragments that covered the entire open reading frame (ORF) were amplified by RT-PCR, and the 3'-untranslated region (UTR) and 5'-UTR were amplified using the 3'-rapid amplification of cDNA ends (RACE) and the 5'-RACE method, respectively. Each of the gene fragments were cloned and sequenced, and whole genome sequences of the strain were obtained by assembling the cDNA fragment sequences. The results showed that the length of the complete genome was 7 380 nucleotides (GenBank accession number: HQ317203). The comparison of nucleotide and deduced amino acid sequences of the isolate was performed against other MNV strains in the GenBank database. A phylogenetic tree based on VP1 nucleotide sequences was constructed using MEGA5.0 software. The homology of nucleotides between the MNV Guangzhou/K162/09/CHN strain and other MNV isolates ranged from 87.4% to 89.7%. Phylogenetic analysis showed that there was a close genetic relationship between the Guangzhou/K162/09/CHN strain and MNV strains isolated from Japan (S7-P2 and S7-PP3 isolates), Korea (K4 isolate), and Germany (Berlin/04/06/DE and Berlin/05/06/DE isolates). This is the first report of the isolation and identification of MNV in China, and the first report of the genetic analysis of its complete genome.
Animals ; Caliciviridae Infections ; veterinary ; virology ; Mice ; Molecular Sequence Data ; Norovirus ; classification ; genetics ; isolation & purification ; physiology ; Open Reading Frames ; Phylogeny ; Rodent Diseases ; virology ; Sequence Homology, Amino Acid ; Viral Proteins ; chemistry ; genetics