OBJECTIVETo construct a cDNA library of osteosarcoma SAOS-2 cells and screen the proteins interacting with the bone morphogenetic protein-2 (BMP-2) to explore the role of BMP-2 in the development of prostate cancer.

METHODSThe total RNA was extracted from SAOS-2 cells, from which poly(A)(+)RNA was purified to generate the cDNA using RT-PCR with primer SMARTIII and primer CDSIIIoligo(dT). The library was constructed and screened by cotransformation of the double-stranded cDNA (dscDNA) and linearized pGADT7-Rec with the bait plasmid to the yeast AH109. The proteins interacting with BMP-2 were screened by AH109 mating with the bait strain Y187, and the interaction was confirmed using far-Western blot analysis.

RESULTSThe cDNA library of human osteosarcoma SAOS-2 cells was constructed, which was characterized by high diversity and sufficient capacity. The dscDNA size range was between 250-5000 bp, with a cotransformation efficiency of 4.3 x 10(5) and recombination efficiency of 1.9 x 10(6), and 4.3 x 10(5) clones were screened. The mating efficiency was 32% and 1.0 x 10(6) clones were screened by the bait of BMP-2, and 4 positive clones was obtained and sequenced.

CONCLUSIONThe diversity and capacity of the cDNA library meet the needs for screening genes related to prostate cancer.

Bone Morphogenetic Protein 2 ; genetics ; Cell Line, Tumor ; Gene Library ; Humans ; Male ; Osteosarcoma ; genetics ; Prostatic Neoplasms ; genetics ; RNA, Neoplasm ; genetics ; Two-Hybrid System Techniques

OBJECTIVETo determine whether testosterone-induced intra-testicular testosterone withdrawal and therefore spermatogenic impairment is associated with looser arrangement of spermatogenic cells in rats.

METHODSAdult male SD rats received intramuscular injection of testosterone undecanoate at 19 mg/(kg x 15 d) for 130 days, and then testicular tissue blocks were obtained for the preparation of methacrylate resin-embedded sections and observation of the changes in testicular histology.

RESULTSApart from such changes as impaired spermiogenesis and spermiation, apparently looser arrangement of spermatogenic cells was seen in 11.5% of the seminiferous tubule profiles, with radial cracks (empty spaces) running towards the tubule lumen being formed between lines, bundles or groups of spermatogenic cells (mainly spermatids and spermatocytes).

CONCLUSIONLooser arrangement of spermatogenic cells is one of the key histological changes resulting from intra-testicular testosterone withdrawal in rats.

Animals ; Male ; Rats ; Rats, Sprague-Dawley ; Seminiferous Tubules ; cytology ; drug effects ; Spermatogenesis ; drug effects ; Testis ; cytology ; drug effects ; pathology ; Testosterone ; administration & dosage ; adverse effects

Adipose-derived stem cells (ASCs) induce therapeutic angiogenesis due to pro-angiogenic cytokines secretion.Superparamagnetic iron oxide (SPIO) nanoparticles are critical for magnetic resonance (MR) tracking of implanted cells.Hypoxia is a powerful stimulus for angiogenic activity of ASCs.In this study,we investigated whether therapeutic potency could be enhanced by implantation of hypoxia-preconditioned SPIO-labeled ASCs (SPIOASCs) into the infarcted myocardium.ASCs and SPIOASCs were cultured under 2％ O2 (hypoxia) or 95％ air (normoxia).Cells were intramyocardially injected into the infarcted myocardium after 48-h culture.We found that hypoxia culture increased the mRNA expression of hypoxia-inducible factor-1 alpha (HIF-lαt) and vascular endothelial growth factor (VEGF) in ASCs and SPIOASCs.The VEGF protein in the conditioned medium was significantly higher in hypoxic ASCs and SPIOASCs than in normoxic ASCs and SPIOASCs.The capillary density and left ventricular contractile function in the infarcted myocardium were significantly higher 4 weeks after implantation with hypoxic ASCs and SPIOASCs than with normoxic ASCs and SPIOASCs.Improvement in the capillary density and left ventricle function didn't differ between hypoxic ASCs-transplanted rats and hypoxic SPIOASCs-transplanted rats.Hypoxic culture enhanced the angiogenic efficiency of ASCs.It was concluded that implantation of hypoxic ASCs or SPIOASCs promotes therapeutic angiogenesis and cardiac function recovery in the infarcted myocardium.SPIO labeling does not impact the beneficial effect of hypoxic ASCs.

OBJECTIVETo compare the efficacy, time to disease progression (TTP), overall survival (OS) and toxicity of FOLFOX6 and TLF regimens for advanced gastric cancer.

METHODSThe clinical data of 81 chemotherapy-naive patients with advanced gastric cancer were analyzed. Of the 81 patients, 41 were treated with FOLFOX6 regimen and 40 with TLF regimen. The patients in FOLFOX6 group received intravenous infusion of L-OHP(100 mg/m2) at day 1, bolus injection of 5-FU (400 mg/m2) at day 1, and continuous intravenous infusion of 5-FU (1200 mg/m2/d) for 22 h at days 1-2, each treatment cycle lasting 14 days. The patients in TCF group received TAX (90 mg/m2) at day 1, bolus injection of 5-FU (400 mg/m2) at days 1-2, and continuous intravenous infusion of 5-FU (400 mg/m2/d) for 22 h at days 1-2, and each treatment cycle also lasted 14 days.

RESULTSThe objective response rates were 48.8% in FOLFOX6 group and 50.0% in TLF group (P=0.962). The median TTP in the two groups was 6.30 months and 6.50 months (P=0.958), with median survival time of 9.80 months and 10.70 months (P=0.578), respectively. The most frequent adverse events were nausea, vomiting and hematologic toxicities. The incidences of grade III-IV leucopenia and neutropenia were lower in FOLFOX6 group than in TLF group, but the difference was not statistically significant (12.2% vs 30.0%, P=0.112; 14.6% vs 32.5%, P=0.126). Three patients in FOLFOX6 group developed intestinal obstruction during the chemotherapy.

CONCLUSIONBoth FOLFOX6 and TLF regimens are effective in treating advanced gastric cancer and the toxicities can be tolerated.

Adenocarcinoma ; drug therapy ; pathology ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Disease Progression ; Female ; Fluorouracil ; adverse effects ; therapeutic use ; Humans ; Leucovorin ; adverse effects ; therapeutic use ; Male ; Middle Aged ; Neoplasm Staging ; Organoplatinum Compounds ; administration & dosage ; adverse effects ; therapeutic use ; Stomach Neoplasms ; drug therapy ; pathology ; Survival Analysis ; Taxoids ; administration & dosage

Adult ; Aged ; Budd-Chiari Syndrome ; diagnostic imaging ; Female ; Humans ; Liver ; diagnostic imaging ; Liver Circulation ; Male ; Middle Aged ; Ultrasonography, Doppler, Color ; Young Adult

OBJECTIVETo design a platform for microarray data analysis and processing in the browser/server mode running in Linux operating system.

METHODSBased on the Apache HTTP server, the platform, programmed with Perl language, integrated R language and Bioconductor packages for processing and analysis of the input data of oligonucleotide arrays and two-color spotted arrays. Users were allowed to submit data and parameter configurations to the platform via the web page, and the results of analysis were also returned via the web page.

RESULTSWith an easy operation and high performance, the platform fulfilled the functions of processing, quality assessment, biological annotation and statistical analysis of the data from oligonucleotide arrays and two-color spotted arrays. Using the platform, we analyzed the gene expression profiles in Mtb-stimulated macrophages of three clinical phenotypes, namely latent TB (LTB), pulmonary (PTB) and meningeal (TBM), and obtained valuable clues for identifying tuberculosis susceptibility genes. We also analyzed the effect of INH treatment on Mycobacterium tuberculosis gene expression in various dormancy models, such as hypoxia and KatG mutant, and found that a set of genes responded to INH treatment during exponential growth but not in dormancy, and that the overall number of differentially regulated genes was reduced in the cells in low metabolic state.

CONCLUSIONThe platform we have constructed integrates comprehensive resources, and with a user-friendly interface, allows direct result visualization to facilitate microarray data analysis.

Computational Biology ; methods ; Databases, Genetic ; Gene Expression Profiling ; methods ; Genetic Predisposition to Disease ; Humans ; Macrophages ; metabolism ; microbiology ; Mycobacterium tuberculosis ; genetics ; Oligonucleotide Array Sequence Analysis ; Software ; Tuberculosis ; genetics ; immunology ; User-Computer Interface

Animals ; Antineoplastic Agents ; administration & dosage ; Chemoembolization, Therapeutic ; methods ; Female ; Hepatic Artery ; Liver Neoplasms, Experimental ; therapy ; Oligopeptides ; administration & dosage ; Rats ; Rats, Wistar

The comparative efficacy of 2 anthelmintics (ivermectin and levamisole) against Baylisascaris transfuga migrating and encapsulated larvae was studied in mice. A total of 60 BALB/c mice inoculated each with about 1,000 embryonated B. transfuga eggs were equally divided into 6 groups (A-F) randomly. Mice of groups A and B were treated with ivermectin and levamisole, respectively, on day 3 post-infection (PI). Mice of groups A-C were killed on day 13 PI. Similarly, groups D and E were treated with ivermectin and levamisole, respectively, on day 14 PI, and all mice of groups D-F were treated on day 24 PI. The groups C and F were controls. Microexamination was conducted to count the larvae recovering from each mouse. The percentages of reduction in the number of migrating larvae recovered from group A (ivermectin) and B (levamisole) were 88.3% and 81.1%, respectively. In addition, the reduction in encapsulated larvae counts achieved by ivermectin (group D) and levamisole (group E) was 75.0% and 49.2%, respectively. The results suggested that, to a certain extent, both anthelmintics appeared to be more effective against migrating larvae than encapsulated larvae. However, in the incipient stage of infection, ivermectin may be more competent than levamisole as a larvicidal drug for B. transfuga.
Animals ; Anthelmintics/*administration & dosage ; Ascaridida Infections/*drug therapy/parasitology ; Ascaridoidea/*drug effects ; Disease Models, Animal ; Female ; Ivermectin/*administration & dosage ; Larva/drug effects ; Levamisole/*administration & dosage ; Male ; Mice ; Mice, Inbred BALB C ; Rodent Diseases/drug therapy/parasitology ; Treatment Outcome

OBJECTIVETo design a versatile genome comparison and visualization platform based on browser/server mode supported by a local server.

METHODSThe server of the platform was Apache HTTP server. Perl was used to integrate such genome alignment package and algorithms as MUMmer, LAGAN, and Mauve for different comparison purposes, and the users could submit data and parameters to the platform via the web page. The results of analysis were also returned via the web page.

RESULTSThe platform could handle multiple data input formats, compare complete and draft genome sequence, alignment pair-wise or multi genome of more divergent species, identify regions of high similarity, locate local nucleotide mutations and large-scale recombination, and display the results by visualization interface. Analysis of the homology of 10 new strains of influenza A virus indicated that PB1 gene might evolve from human H3N2 viruses, PB2 and PA genes from avian H3N2 viruses, and HA and NS genes from swine H1N1 viruses. Alignment of Mycobacterium tuberculosis (H37Rv, CDC1551) and Mycobacterium bovis (AF2122/97) genomes revealed that sequence insertion/deletion and duplication were the major source of genomic differences.

CONCLUSIONThe platform integrate comprehensive resources with a user-friendly interface and intuitive result visualization to facilitate conventional study of comparative genomics.

Base Sequence ; Comparative Genomic Hybridization ; methods ; Genome ; Genomics ; methods ; Influenza A Virus, H1N1 Subtype ; genetics ; Influenza A Virus, H3N2 Subtype ; genetics ; Molecular Sequence Data ; Mycobacterium tuberculosis ; genetics ; Software ; User-Computer Interface

A GeXP based multiplex PCR assay was developed to simultaneously detect six different chicken respiratory viruses including H5, H7, H9 subtypes of avian influenza virus(AIV), new castle disease virus (NDV), infectious bronchitis virus(IBV) and infectious laryngotracheitis virus(ILTV). According to the conserved sequences of genes of each pathogen, seven pairs of specific primers were designed, and the reaction conditions were optimized. The specificity and accuracy of GeXP were examined using samples of single and mixed infections of virus. The sensitivity was evaluated by performing the assay on serial 10-fold dilutions of cloned plasmids. To further evaluate the reliability, thirty-four clinical samples were detected by GeXP. The corresponding specific fragments of genes were amplified. The detection limit of GeXP was 10(2) copies/microL when all of 7 pre-mixed plasmids containing target genes of six chicken respiratory viruses were present. In the detection of thirty-four clinical samples, the results of GeXP were accorded with the viral isolation completely. In conclusion, this GeXP assay is a rapid, specific, sensitive and high-throughput method for the detection of chicken respiratory virus infections. It can be applied in rapid differential diagnosis for clinical samples, and also provide an effective tool to prevent and control chicken respiratory diseases with similar clinical symptoms.
Animals ; Chickens ; Influenza A virus ; classification ; genetics ; isolation & purification ; physiology ; Influenza in Birds ; diagnosis ; virology ; Multiplex Polymerase Chain Reaction ; methods ; Poultry Diseases ; diagnosis ; virology ; Respiratory Tract Infections ; diagnosis ; veterinary ; virology