[Therapeutic effect of adenovirus-mediated apoptin gene transfer combined with ADM and CDDP on hepatocellular carcinoma in mice].

Objective: To study the therapeutic effect adenovirus-mediated apoptin gene transfer combined with ADM and CDDP on hepatocellular carcinoma in mice.

Methods: In c57BL/ 6 mice bearing hepatocellular carcinoma, the changes of tumor volume, histomorphology, tumor inhibition rate and the side effects were observed after intratumoral injection of adenovirus containing apoptin gene and ADM and CDDP.

Results: Seven days after the treatment, the mean volume of the tumor in the mice receiving intratumoral apoptin-containing adenovirus injection combined with ADM and CDDP reduced significantly as compared with that in mice treated with adenovirus vehicle and control group.

Mitochondrial DNA from colorectal cancer cells promotes the malignant phenotype of NIH3T3 cells.

We investigated the potential role of mitochondrial DNA (mtDNA) in colorectal carcinogenesis by constructing a eukaryotic expression vector of the mitochondrial D-loop gene from colorectal cancer cell SW480 and transfected NIH3T3 cells. The NIH3T3/SW480 cells exhibited a significantly increased growth rate and colony formation rate, and also had a decreased apoptotic rate. Polyploidy and aberrant chromosomes were detected in the NIH3T3/SW480 cells by chromosome karyotype analysis.

[Spheres isolated from Colo205 cell line possess cancer stem-like cells under serum-free culture condition].

Objective: Isolation and expansion tumor spheres from colorectal cancer cell line Colo205 cultured in serum-free medium(SFM) supplemented with human recombinant EGF and bFGF.

Methods: Colo205 cells were cultivated in SFM,while cells cultivated in serum-supplemented medium(SSM) served as the control. Cells morphology were observed by optical microscope, and expression of intestinal stem cells marker Musashi-1 was detected by immunocytochemical.

[Peripheral blood p53 single nucleotide polymorphism analysis for early diagnosis of colorectal carcinoma].

Objective: To evaluate the role of p53 gene mutation in colorectal carcinoma and assess the value of peripheral blood p53 single nucleotide polymorphism (SNP) in early diagnosis of colorectal carcinoma.

Methods: NSP in axons 5-8 of p53 gene was detected using ligase detection reaction-polymerase chain reaction (LDR-PCR) in the peripheral blood of 100 patients with colorectal cancer and 100 healthy subjects.

Results: The mutation rate of p53 gene was 24% (24/120) in colorectal carcinoma patients and 0% (0/120) in the healthy subjects (P<0.

[Effects of angiotensin II and aldosterone on NF-kappaB binding activity in hepatic stellate cells].

Objective: To investigate the signal transduction mechanism underlying the effects of angiotensin II (AngII) and aldosterone (Aldo) on nuclear factor-kappaB (NF-kappaB) DNA binding activity.

Methods: Sixty male Wistar rats were randomly divided into 4 groups: model group (Mo group), injected with CCl(4) subcutaneously twice a week to establish a model of hepatic fibrosis; perindopril group (Pe group), injected with CCl(4) subcutaneously twice a week and perfused with perindopril once a day; losartan group (Lo group), injected with CCl(4) subcutaneously twice a week and perfused with losartan once a day; and control group (Nc group), injected with olive oil subcutaneously. The rats were killed in batches respectively 4 and 6 weeks after and their livers were collected to undergo Masson staining and be observed by light microscope.

[Effects of combined use of curcumin and catechin on cyclooxygenase-2 mRNA expression in dimethylhydrazine-induced rat colon carcinogenesis].

Objective: To examine the effect of combined use of curcumin and catechin on the number of aberrant crypt foci (ACF) and expression levels of cyclooxygenase-2 (COX-2) mRNA in rat colon carcinogenesis. Methods Dimethylhydrazine (DMH)-induced rats colon carcinogenesis model was used for evaluation of the synergistic inhibitory effect between curcumin and catechin in light of ACF formation and tumor incidence. COX-2 mRNA expression was also detected in rat colon carcinogenesis.

[Modulation of intestinal mucosal inflammatory factors by curcumin in rats with colitis].

Objective: To explore the mechanism of modulation of intestinal mucosal inflammatory factors by curcumin, the inhibitor of the transcriptional factor nuclear factor -kappaB (NF-kappaB), in rats with trinitrobenzene sulfonic acid (TNBS)-induced colitis, and screen for a targeted therapeutic agent for treatment of inflammatory bowel disease (IBD).

Methods: Rats with TNBS-induced colitis were fed with diet containing 2.0% curcumin (treatment group), 0.

[Screening of differentially expressed genes related to laterally spreading tumor by cDNA microarray].

Objective: To screen differentially expressed genes between laterally spreading tumor (LST) cell line and common colon carcinoma cell lines, and identify new targets and strategies for exploring the pathogenesis of colorectal tumor.

Methods: The total RNA was extracted from the LST, SW480 and LoVo cells, from which purified mRNAs were obtained. The PCR products of 18 816 genes were blotted onto a fibrous membrane to generate the microarray.

Competitive inhibition of adherence of enterotoxigenic Escherichia coli, enteropathogenic Escherichia coli and Clostridium difficile to intestinal epithelial cell line Lovo by purified adhesin of Bifidobacterium adolescentis 1027.

Aim: To observe competitive inhibition of adherence of enterotoxigenic Escherichia coli (ETEC), enteropathogenic Escherichia coli (EPEC) and Clostridium difficile (C. difficile) to intestinal epithelial cell line Lovo by purified adhesin of Bifidobacterium adolescentis 1027 (B. ado 1027).

[Effect of Bifidobacterial adhesin on lipopolysaccharide- and H2O2-induced proliferation and apoptosis of intestinal epithelial cells in vitro].

Objective: To study the effect of Bifidobacterial adhesin on proliferation and apoptosis of intestinal epithelial cell induced by lypopolysaccharide (LPS) and H(2)O(2) in vitro.

Methods: With (3)H-TdR incorporation method, flow cytometry and fluorochrome staining, the proliferation and apoptosis of intestinal epithelial cells induced by LPS and H(2)O(2) in vitro were studied.

Results: LPS at the dose of 100 microg/L effectively stimulated the proliferation and apoptosis of cells, whereas H(2)O(2) at the dose of 200 micromol/L obviously restrained the proliferative ability while enhanced the apoptosis of the cells.

[Electron microscopic observation of primary cultured laterally spreading tumor cell line].

Objective: To observe the microscopic characteristics of laterally spreading tumor (LST) cell line in primary culture.

Methods: The cells isolated from a rectum LST specimen obtained by endoscopic mucosal resection was primary cultured, followed by observation with scanning and transmission electron microscope in comparison with the cells of adenocarcinoma and normal mucosa of the rectum.

Results: Scanning and transmission electron microscopes both revealed numerous microvilli covering the surface of the LST cells, and the cytoplasm contained large quantity of lysosomes, mitochondria and phagosomes.

[Contribution of eukaryotic initiation factor-4E inhibition to heparanase expression and activity in human colon adenocarcinoma cell line LS-174T].

Objective: To determine whether the eukaryotic initiation factor-4E (eIF-4E) is involved in the cap-dependent translational regulation of heparanase and study the correlation between heparanase expression and metastatic potential of LS-174T cells.

Methods: The protein and mRNA levels of inhibited eIF-4E were tested by Western blot and RT-PCR. Heparanase activity was defined as the ability to degrade high molecular weight (40-100 000) radiolabeled ((35)S) heparan sulfate (HS) substrate into low molecular weight (5-15 000) HS fragments.

[Expression of pro-inflammation cytokines and activation of nuclear factor kappab in the intestinal mucosa of mice with ulcerative colitis].

Objective: To investigate the expression of pro-inflammation cytokines and activation of nuclear factor kappaB (NF-kappaB) in mouse models of ulcerative colitis.

Methods: Mouse models of ulcerative colitis were established by oral administration of 5% dextran sulfate sodium for 7 d, and the expression of tumor necrosis factor (TNF)- alpha and interleukin (IL)-1beta in the intestinal mucosa were detected by semi-quantitative reverse transcriptional (RT) PCR. The activation of NF-kappaB in the intestinal mucosa was evaluated by electrophoretic mobility shift assay (EMSA).

[The effects of bifidobacterium on the intestinal mucosa of the patients with ulcerative colitis].

Objective: To investigate the effects of bifidobacterium (Bf) on the intestinal mucosa of the patients with ulcerative colitis (UC).

Methods: Thirty patients in clinical and endoscopic remission by sulphasalazine and glucocorticoid were randomized to receive either Bifid Triple Viable capsule (BIFICO), 420 mg/d, or an identical placebo (starch) for 8 weeks. Fecal samples were collected for stool culture before and after treatment.

Contribution of eIF-4E inhibition to the expression and activity of heparanase in human colon adenocarcinoma cell line: LS-174T.

Aim: Heparanase degrades heparan sulfate proteoglycans (HSPGs) and is a critical mediator of tumor metastasis and angiogenesis. Recently, it has been cloned as a single gene family and found to be a potential target for antimetastasis drugs. However, the molecular basis for the regulation of heparanase expression is still not quite clear.

[A novel method for fabrication of tissue microarray].

Background & Objective: Tissue chip (tissue microarray, TMA) is one of the most important biochip techniques, just following the gene chip and the protein chip, which is one of the most important functional genomics and proteomics research methods in the post-genomic era. However, the present TMA technology has certain shortcomings, such as lack of advanced instruments, tedious procedure, and low sampling accuracy, etc. This new method was designed to improve the technology of TMA.

Effect of eIF-4E inhibition on heparanase mRNA and its expression in colon adenocarcinoma cell.

Objective: To verify whether inhibition of the overexpressed eukaryotic initiation factor-4E (eIF-4E) in human colon adenocarcinoma cell line LS-174T may facilitate the degradation of heparanase mRNA and alter the translation and expression levels of heparanase protein.

Methods: A 20-mer antisense s-oligodeoxynucleotide (asODN) targeted against the translation start site of eIF-4E mRNA was introduced into LS-174T cells by means of lipid-mediated DNA-transfection, followed by Western blotting analysis and reverse transcription-PCR to determine eIF-4E protein and mRNA levels, respectively. Northern methods was applied to determine heparanase mRNA expression level, with the alterations of heparanase expression assessed by Western blotting analysis.

[Construction and identification of the cDNA phage expression library for human colorectal cancer antigens].

Objective: To construct a cDNA phage expression library for human colorectal carcinoma antigens.

Methods: After the total RNA was extracted from human colorectal cancer tissues, the single-strand and double-strand cDNA were synthesized through reverse transcriptase PCR and long-distance PCR, with the cDNA fragments smaller than 500 bp removed and the remaining cDNA combined with the right and left arms of dephosphorylated lambdaTriplEx2 phage vector. The recombinant phage were then packaged in vitro by MaxPlax Packaging extract, and a small portion of the packaged phage was used to infect E.

Pleckstrin homology domain of G protein-coupled receptor kinase-2 binds to PKC and affects the activity of PKC kinase.

Aim: To study the detail mechanism of interaction between PKC and GRK(2) and the effect of GRK(2) on activity of PKC.

Methods: The cDNA of pleckstrin homology (PH) domain located in GRK(2) residue 548 to 660 was amplified by PCR with the mRNA of human GRK(2) (beta1-adrenergic receptor kinase) as template isolated from human fresh placenta, the expression vector pGEX-PH inserted with the aboved cDNA sequence for GRK(2) PH domain protein and the expression vectors for GST (glutathion-s-transferase) -GRK(2) PH domain fusion protein, BTK (Bruton's tyrosine kinase) PH domain and GST protein were constructed. The expression of GRK(2) in culture mammalian cells (6 cell lines: PC-3, MDCK, SGC7901, Jurkat cell etc.