Objective To use bioinformatics methods to analyze large amounts of data generated by gene chips and to screen common key genes in hepatocellular carcinoma in human and rat.Methods For search of the medical literature,3 sets of gene chip with data which met our predetermined criteria were downloaded from the GEO database.The data were standardized by using the bioconductor and R version of the 2.10.1 version.The original data of the affymetrix platform were normalized with background correction,standardized and transformed into log2 by using the algorithm of the affy packages RMA.The TTEST function of the excel was then used to calculate the significance of each gene.The DAVID was used for gene ID conversion and a table was established for samples and the corresponding gene expression data.A meta analysis was performed to calculate the common genes of human and rat.An enrichment regulation pathway was gained with the KEGG in the DAVID library. Results There were 26 common expression genes in the development process of hepatocellular carcinoma in human and rat.Five of these genes were up-regulation genes,while twenty-one were down-regulation genes.An enrichment pathway,which is a focal adhesion pathway,was found and this pathway has been reported to be associated with development of hepatocellular carcinoma.Conclusion With bioinformatics,we were able to screen common key genes and a pathway which were closely related to development of hepatocellular carcinoma in human and rat.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Immunoglobulins, Intravenous ; therapeutic use ; Infant ; Infant, Newborn ; Male ; Prednisone ; therapeutic use ; Purpura, Thrombocytopenic, Idiopathic ; drug therapy

Objective To evaluate and summarize a new way of forward intensity modulated radia- tion therapy (IMRT) plan for nasopharyngeal carcinoma with 3Dmulti-leaf collimator (MLC) planning sys- tems is practised routinely in our department.Methods From September 2000 to July 2003 and November 2005 to March 2006,78 patients with nasopharyngeal carcinoma who were treated with eonformal radiothera- py used CT simulation for localizing,then doctors supervised the delineation of the planned tumor volume (PTV),gross tumor volume (GTV) and other sensitive organs on ACQsim workstation,and then sent the CT imagines to ELEKTA Precise Plan by DICOM RT Ethernet.Then,the physicists take over the responsibility of all directions of beam projection according to these organs and PTV shape,and completed the IMRT plan by manual correction making and the optimization of fields or segments shaped with forward 3D planning sys- tem.Results To analyse all the patient's dose distributions taking conformal radiotherapy in our hospital, we found the dose distribution and DVH data excellent,even better than the inverse planning of nasopharyn- geal carcinoma.Conclusions Intensity modulated radiation therapy (IMRT) is an advanced radiotherapy technique.A very good IMRT plan of nasopharyngeal carcinoma by forward planning can be obtained with 3D multiple leaf collimator (MLC) planning system.This planning method is more flexible,but the radia- tion physicists should be very much experienced in professional skill.

Objective To investigate the mechanism of hypoxia regulate osteopontin (OPN) secreting by mature dendritic cells (mDCs). Methods CD14 + cells were enriched using anti-CD14 immunomagnetic beads, for inducing to mDCs, CD14 + cells were cultured with GM-CSF and IL-4 in hypoxia or normoxiain vitro. Concentration of OPN and TGF-β1 in supernatant were detected by sandwich ELISA, OPN mRNA detected by RT-PCR. Approach regulating function of A2 R in expressing of OPN by mDCs by using NECA (surrogate of adenosine), A2R agonist (CGS21680), A2R antagonist (SCH58261) and investigate role of TGF-β1 in this process by using rhTGF-β1 and anti-TGF-β1 Ab. Results Hypoxia inreased the level of OPN and OPN mRNA in mDCs, and this effect could be reversed by A2 R antagonist. Under normoxia,both NECA and A2R agonist (CGS21680) could upregulate the level of OPN and OPN mRNA in mDCs significantly, but this positive effect could be reversed by A2 R antagonist. A2 R played a role in regulating TGF-β1, and confirmed TGF-β1 involved in regulation of OPN by using rhTGF-β1 and anti-TGF-β1 Ab. Conclusion High adenosine induce the generation of TGF-β1 through the A2R on mDCs, and then TGF-β1 raise the OPN secreting by mDCs.

OBJECTIVETo establish discriminant functions of diarrhea-predominant irritable bowel syndrome (IBS-D) by studying it from quantitative diagnosis angle, hoping to reduce interference of subjective factors in diagnosing and differentially diagnosing Chinese medical syndromes of IBS-D.

METHODSA Chinese medical clinical epidemiological survey was carried out in 439 IBS-D patients using Clinical Information Collection Table of IBS. Initial syndromes were obtained by cluster analysis. They were analyzed using step-by-step discrimination by taking information of four Chinese medical diagnostic methods and serum brain-gut peptides (BGP) as variables.

RESULTSClustering results were Gan stagnation Pi deficiency syndrome (GSPDS), Pi-Wei weakness syndrome (PWWS), Gan stagnation qi stasis syndrome (GSQSS), Pi-Shen yang deficiency syndrome (PSYDS), Pi-Wei damp-heat syndrome (PWDHS), cold-damp disturbing Pi syndrome (CDDPS). Of them, GSPDS was mostly often seen with effective percentage of 34. 2%, while CDDPS was the least often seen with effective percentage of 5.5%. A total of 5 discriminant functions for GSPDS, PWWS, GSQSS, PSYDS, and PWDHS were obtained by step-by-step dis- crimination method. The retrospective misjudgment rate was 4.1% (16/390), while the cross-validation misjudgment rate was 15.4% (60/390).

CONCLUSIONThe establishment of discriminant functions is of value in objectively diagnosing and differentially diagnosing Chinese medical syndromes of IBS-D.

Alarmins ; Brain ; Cluster Analysis ; Diarrhea ; classification ; diagnosis ; Hot Temperature ; Humans ; Irritable Bowel Syndrome ; classification ; diagnosis ; Medicine, Chinese Traditional ; Qi ; Retrospective Studies ; Surveys and Questionnaires ; Yang Deficiency

Antioxidants ; pharmacology ; Ascorbic Acid ; pharmacology ; Carbon Disulfide ; adverse effects ; antagonists & inhibitors ; Cells, Cultured ; Endothelium, Vascular ; drug effects ; Humans ; Umbilical Veins ; cytology ; drug effects ; metabolism

Objective To retrospectively analyze the therapeutic effect on patients with tonsillar carcinoma and factors affecting their prognosi. Methods Clinical data of 61 patients pathologically confirmed with tonsillar carcinoma without distant metastasis were analyzed. All the patients were treated in Cancer Center of Sun Yat-sen University from April 1997 to April 2008. There were 2 patients with undifferentiated carcinoma, 26 with poorly differentiated squamous cell carcinoma and 33 with median-well differentiated squamous cell carcinoma. According to the AJCC 2002 staging criteria for head-neck cancers, there were 9 staged Ⅰ cases, 7 staged Ⅱ cases,23 staged Ⅲ cases and 22 staged Ⅳ cases. The treatment was radiotherapy alone in 27 cases, radiotherapy combined with chemotherapy in 23 cases, surgery combined with postoperative radiotherapy in 6 cases, neoadjuvant chemotherapy plus surgery combined with postoperative radiotherapy in 3 cases, radiotherapy with salvage surgery in 2 cases. Results The overall 5-year survival rate was 50. 2%. For 16 cases with staged Ⅰ-Ⅱ staged, there were 8 cases with radiotherapy alone, 5 years survival was 50. 0%, 6 cases with surgery combined with postoperative radiotherapy, 5 years survival was 83.3%. The difference between the two treatments was not significant in statistics (P=0.318). For Ⅲ-Ⅳ staged 45 cases, there were 19 cases with simple radiotherapy, 5 years survival was 51.5%, 21 cases with radiotherapy combined with chemotherapy, 5 years survival was 36.4%, 5 cases with surgery combined with postoperative radiotherapy, 5 years survival was 75.0%. The difference among the three treatments was not significant in statistics (P=0.239). According to T stages, the 5-year survival rates of stage T1-T4 cases were 91.8%, 46.8%, 29.1%, 0% respectively (χ~2=30.168, P<0.001). Multivariate analysis demonstrated that T stage, therapeutic effect of primary site and cervical metastatic lymph node were the independent prognostic factors (P<0.05). Conclusions T stage, the therapeutic effect of primary site and cervical metastatic lymph node were the independent prognostic factors. For Ⅰ-Ⅱ staged tonsillar tumor cases, based on organ preservation, were tendency to choice simple radiotherapy. For Ⅲ-Ⅳ staged cases,yet the ralationships between therapeutic mode and therapeutic effect still need further researches.

Objective:To study the acidic heteropolysac charides of Cynomorium songaricum.Method:The polysaccharides were purified by Sephadex G-100 and G-150 gel column chromatography.Purity and molecular weight of the polysaccharides were determined by high performance gel permeation chromatography; neutral sugars composition were identified by PC, TLC and GC. Uronic acids were determined by carbazole method.Result:The molecular weight of SYP-A and SYP-B were estimated to be 3.1×105 and 2.8×105 respectively, and neutral sugars were composed of galactose, glucose, arabinose, rhamnose, mannose and ribose. The molar ratio for SYP-A and SYP-B were 5.1∶4.1∶1.6∶1.0∶0.5∶0.3 and 5.2∶4.2∶1.5∶1.0∶0.5∶0.2 respectively, The contents of uronic acids were 10.7% and 10.5% respectively.Conclusion:SYP-A and SYP-B are homogeneous acidic heteropolysaccharides.

OBJECTIVETo express human osteoprotegerin (OPG) in E. Coli and analyze its bioactivity in vitro.

METHODSSynthetic oligonucleotides were used to amplify human OPG gene by RT-PCR from total RNA of human osteosarcoma cell line MG63. The OPG cDNA coding for 380 amino acid residues was inserted into prokaryotic expression vector pRSET-A, transformed into competent E. Coli BL21, and induced by 0.1 mmol/l IPTG. SDS-PAGE and Western blot were performed to identify OPG-6His fusion protein. After purified by affinity chromatography, 1,000 microg/L or 1,500 microg/L of OPG-6His were added into the mouse bone marrow cells culture medium. The number of tartrate-resistant acid phophatase (TRAP)-positive multinucleated cells and resorption pits were counted to assess the bioactivity of expression products.

RESULTSThe sequence of OPG mature peptide encoding cDNA obtained in this experiment was as same as reported. SDS-PAGE showed 24% of total bacterial protein was of OPG-6His fusion protein. Western blot assay demonstrated that the molecular weight of recombinant protein was about 46 KD and could react specifically with human anti-OPG antibody. The mouse bone marrow cells were induced by 1alpha, 25-dihydroxyvitaminD3 (10(-8) mol/L) and Dexamethasone (10(-7) mol/L) to form osteoclastic-like multinucleated cells. 1,500 microg/L of purified OPG-6His protein could decrease the number of resorption pits and TRAP-positive multinucleated cells in vitro (P < 0.05), but it didn't show the same effects when the concentration of OPG-6His fusion protein was of 1,000 microg/L.

CONCLUSIONSHuman OPG-6His fusion protein is expressed and purified in E. Coli. The expression products have moderate inhibitory effects on osteoclast differentiation and bone resorption in vitro only when excessive amount of proteins are added into the culture medium, indicating that prokaryotic expression of fuctionalal OPG protein awaits further investigation.

Cell Differentiation ; drug effects ; Cell Line, Tumor ; Cloning, Molecular ; Escherichia coli ; genetics ; Glycoproteins ; biosynthesis ; genetics ; Humans ; Osteoclasts ; drug effects ; physiology ; Osteoprotegerin ; Receptors, Cytoplasmic and Nuclear ; biosynthesis ; genetics ; Receptors, Tumor Necrosis Factor ; Recombinant Fusion Proteins ; biosynthesis ; pharmacology

Using the isolated total RNA from osteosacoma cell line MG63, the cDNA encoding human OPG was amplified by RT-PCR. A recombinant adenoviral vector carrying cDNA of OPG was constructed and OPG expression in mouse myoblast C2C12 cells was confirmed by Western blot and ELISA. The secreted expression of OPG protein persisted more than 6 weeks in vitro, and the growth of C2C12 cells infected by recombinant adenoviral were in good state. Osteoclasts derived from mouse bone marrow cells infected with recombinant adenoviral made less number of TRAP positive cells and resorption pits formed on dentine slices.
Adenoviridae ; genetics ; Animals ; Blotting, Western ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; Genetic Vectors ; genetics ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Osteoclasts ; metabolism ; Osteoprotegerin ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction