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.

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 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.

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

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

BACKGROUND/AIMS: The relationship between C-peptide levels and gastrointestinal (GI) symptoms in type 2 diabetic patients is not clear. The purpose of this study is to examine the association between fasting C-peptide and GI symptoms of gastroparesis in type 2 diabetic patients. METHODS: We recruited 333 type 2 diabetic patients into the present study. All patients filled out questionnaires of gastroparesis cardinal symptom index (GCSI) to evaluate GI symptoms. Hospital anxiety and depression scale were adopted to define anxiety and depression. Patients with GCSI scores ≥ 1.9 were regarded as having symptoms of gastroparesis. RESULTS: In our study, 71 (21.3%) type 2 diabetic patients had GCSI scores ≥ 1.9. In comparison to patients with scores < 1.9, those with scores ≥ 1.9 had significantly lower fasting c-peptide levels (1.49 ng/mL vs 1.94 ng/mL, P < 0.001), higher prevalence of depression (40.9% vs 18.3%, P < 0.001) and anxiety (28.2% vs 13.0%, P = 0.002). Multivariate logistic regression revealed that fasting C-peptide was still significantly associated with symptoms of gastroparesis (odds ratio, 0.67; 95% confidence intervals, 0.48–0.94; P = 0.021), even after adjustments for age, sex, body mass index, HbA1c, current smoking and drinking status, anxiety, and depression. Furthermore, linear regressions showed that fasting C-peptide was independently and negatively related to GCSI scores (standardized regression coefficient, −0.29; P < 0.001) in patients with at least one GI symptom. CONCLUSION: GI symptoms of diabetic gastroparesis affect approximately 20% of type 2 diabetes patients and are associated with lower fasting C-peptide levels independent of depression and anxiety status.
Anxiety ; Anxiety Disorders ; Body Mass Index ; C-Peptide* ; Depression ; Depressive Disorder ; Diabetes Mellitus ; Drinking ; Fasting* ; Gastroparesis* ; Humans ; Linear Models ; Logistic Models ; Prevalence ; Smoke ; Smoking

OBJECTIVETo investigate the effects of overexpression of exogenous Notch1 in human tongue squamous cell carcinoma (TSCC) cells on cell growth and expression of epidermal growth factor receptor (EGFR) in vitro.

METHODSHuman TSCC cell line Tca8113 cells were transiently transfected with the eukaryotic expression plasmid pRAMIC-IRES2-EGFP encoding exogenous intracellular fragment of Notch1 and control plasmid pIRES2-EGFP by Lipofectamine 2000, respectively. Untransfected parental Tca8113 cells served as control. The cell proliferation was evaluated by methyl thiazolyl tetrazolium(MTT) assay. The apoptosis was assessed by flow cytometry. The mRNA and protein levels of Notch1 and EGFR in Tca8113 cells were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot. The expression of EGFR protein in Tca8113 cells was detected by immunocytochemistry.

RESULTSMTT assay showed that the cell proliferation of Tca8113 cells transfected with pRAMIC-IRES2-EGFP was significantly inhibited as compared with controls (P < 0.05). After transfected with pRAMIC-IRES2-EGFP for 48 h, the apoptosis rate of Tca8113 cells was significantly higher than those of Tca8113 cells transfected with pIRES2-EGFP and untransfected Tca8113 cells (P < 0.05), and Notch1 expression was significantly increased at mRNA (P < 0.05) and protein (P < 0.05) levels, while EGFR expression was significantly decreased at mRNA (P < 0.05) and protein (P < 0.05) levels.

CONCLUSIONOverexpression of exogenous Notch1 may inhibit cell growth and down-regulate EGFR expression in TSCC cells.

Apoptosis ; Carcinoma, Squamous Cell ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; Humans ; In Vitro Techniques ; RNA, Messenger ; Receptor, Epidermal Growth Factor ; Tongue Neoplasms ; Transfection