[Clinical research of patients with acute or chronic hepatic failure treated with molecular adsorbent recirculating system].

Objective: To summarize the experience of a single treatment using molecular adsorbent recirculating system (MARS) in patients with acute-on-chronic liver failure.

Methods: Twenty-five eases treated by MARS-artificial liver were followed up and reviewed.

Results: The levels of serum total bilirubin, non-conjugated bilirubin and blood ammonia were significantly decreased from (618.

Expression and function of classical protein kinase C isoenzymes in gastric cancer cell line and its drug-resistant sublines.

Aim: To investigate the expression and function of classical protein kinase C (PKC) isoenzymes in inducing MDR phenotype in gastric cancer cells.

Methods: Two cell lines were used in the study: gastric cancer cell SGC7901 and its drug-resistant cell SGC7901/VCR stepwise-selected by vincristine 0.3, 0.

[Development of oral DNA vaccine based on MG(7)-Ag mimotope of gastric cancer].

Objective: To develop an oral DNA vaccine based on MG(7)-Ag mimotope of gastric cancer using attenuated Salmonella typhimurium and evaluate its efficacy and protective effect.

Methods: The eukaryotic expression vector including the MG(7)-Ag mimotope and a Th epitope was constructed, and then transduced into an attenuated Salmonella typhimurium to get the oral DNA vaccine. C57BL/6 J mice were orally immunized with 1 x 10(8) cfu Salmonella transfectants, with Salmonella harboring empty plasmid, with phophate buffered saline (PBS) as control.

[Local cytokines profile in gastric cancer lesions].

Objective: To study the characteristics of local cytokines profile in human gastric cancer lesions and offer guides for immunotherapy of gastric cancer.

Methods: The cytokines profile in isolated CD4+, CD8+ T subsets and epithelial cells from cancer tissue, contrasted with benign gastric mucosa of 15 gastric cancer patients were analyzed by a highly sensitive radioactivity labeled semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) technique.

Results: The predominance of increased Th2 and reduced Th1 immune response in both CD4+ and CD8+ T subsets from cancer tissue, compared with benign gastric mucosa, were observed.

[The effects of C2 gene on growth of gastric cancer cells].

Objective: To investigate the effect of a newly cloned full length gene, C2 gene, which encodes human eukaryotic translation initiation factor, on growth of gastric cancer cells.

Methods: A constructed eukaryotic vector carrying the full length of C2 gene was amplified, purified and transfected into gastric cancer cell line-SGC 7901. Expression of C2 protein was examined with fluorescence activated cell sorting (FACS) and immunohistochemical staining.

Effect of L-NAME on nitric oxide and gastrointestinal motility alterations in cirrhotic rats.

Aim: To investigate the effect of L-NAME on nitric oxide and gastrointestinal motility alterations in cirrhotic rats.

Methods: Rats with cirrhosis induced by carbon tetrachloride were randomly divided into two groups, one n =13 receiving 0.5mg.

Expression and identification of recombinant soluble single-chain variable fragment of monoclonal antibody MC3.

Aim: To generate soluble single chain variable fragments (ScFv) of monoclonal antibody MC3 recognizing colorectal and gastric carcinomas.

Methods: mRNA was isolated from the hybridoma cell line producing MC3 and the DNAs encoding variable domains of heavy and light chains (VH and VL) of the antibody were amplified separately by RT-PCR and assembled into ScFv DNA with a linker DNA. The ScFv DNA was ligated into the phagemid vector pCANTAB5E and the ligated sample was transformed into E.

Direct effect of croton oil on intestinal epithelial cells and colonic smooth muscle cells.

Aim: To investigate the direct effect of croton oil (CO) on human intestinal epithelial cell (HIEC) and guinea pig colonic smooth muscle cells in vitro.

Methods: Growth curves of HIEC were drawn by MTT colorimetry. The dynamics of cell proliferation was analyzed with flow cytometry, and morphological changes were observed under light and electron microscopy after long-term (6 weeks) treatment with CO.

Expression and bioactivity identification of soluble MG7 scFv.

Aim: To examine the molecular mass and identify the bioactivity of MG7 scFv for its application as a targeting mediator in gene therapy of gastric cancer.

Methods: Two strongly positive recombinant phage clones screened from MG7 recombinant phage antibody library were separately transfected into E.coli TG1.

Differential display of vincristine-resistance-related genes in gastric cancer SGC7901 cell.

Aim: To isolate and clone the vincristine-resistance-related genes in gastric cancer SGC7901 cell line and to clarify the multidrug-resistant molecular mechanism of gastric cancer cells.

Methods: The modified differential-display polymerase chain reaction (DD-PCR) was used to examine differences in the mRNA composition of Vincristine-resistant gastric cancer SGC 7901 cells (SGC7901/VCR), induced by vincristine sulfate versus SGC7901cells. The differentially expressed cDNA fragments were confirmed by reverse Northern analysis, sequencing, BLAST analysis and Northern bolt analysis.

Suppression of P-gp induced multiple drug resistance in a drug resistant gastric cancer cell line by overexpression of Fas.

AIM:To observe the drug sensitizing effect and related mechanisms of fas gene transduction on human drug-resistant gastric cancer cell SGC7901/VCR (resistant to Vincristine).METHODS:The cell cycle alteration was observed by FACS. The sensitivity of gastric cancer cells to apoptosis was determined by in vitro apoptosis assay.

Cloning and identification of an angiostatic molecule IP10/crg -2.

AIM:To obtain human and murine cDNAs encoding IFN-gamma inducible protein 10 (IP-10) and cytokine responsive gene-2 (Crg-2).METHODS:The encoding genes of IP-10 and Crg-2 were amplified by RT-PCR from cultured human fibroblast cells and Balb/c mouse liver treated by IFN-gamma and TNF-alpha,respectively, and cloned into plasmids of pUC19 and pGEM3Zf(+).RESULTS:The nucleotide sequences of the amplified DNA were confirmed by endonucleases digestion and sequencing.

Down-regulation of Hsp90 could change cell cycle distribution and increase drug sensitivity of tumor cells.

AIM:To construct Hsp90 antisense RNA eukaryotic expression vector, transfect it into SGC7901 and SGC7901/VCR of MDR-type human gastric cancer cell lines, HCC7402 of human hepatic cancer and Ec109 of human esophageal cancer cell lines, and to study the cell cycle distribution of the gene transected cells and their response to chemotherapeutic drugs.METHODS:A 1.03kb cDNA sequence of Hsp90beta was obtained from the primary plasmid phHSP90 by EcoR I and BamH I nuclease digestion and was cloned to the EcoR I and BamH I site of the pcDNA by T4DNA ligase and an antisense orientation of Hsp90beta expression vector was constructed.

Antisense to cyclin D1 reverses the transformed phenotype of human gastric cancer cells.

AIM:To further investigate the effect of cyclin D1 on the biologic behavior of cancer cells and its potential role in gene therapy of tumor.METHODS:A cyclin D1 subcloning plasmid termed BKSD1 was constructed by subcloning the human cyclin D1 cDNA into Bluescript-KS, a plasmid vector with a pair of T7 and T3 promoters, with recombinant DNA technology of molecular biology. So,it is easy to generate digoxigenin (DIG)-labeled RNA probes of antisense and sense to cyclin D1 using RKSD1 as a template vector.

Transduction of Fas gene or Bcl-2 antisense RNA sensitizes cultured drug resistant gastric cancer cells to chemotherapeutic drugs.

AIM:To compare the expression level of Fas gene and Bcl-2 gene in gastric cancer cells SGC7901 and gastric cancer MDR (multidrug resistant) cells SGC7901/VCR,to transduce Fas cDNA and Bcl-2 antisense nucleic acid into SGC7901/VCR cells respectively, and to observe the expression of two genes in transfectants and non-transfectants as well as their drug sensitivity.METHODS:Eukaryotic expression vector pBK-Fas cDNA and pDOR-anti Bcl-2 were constructed and transfected into SGC7901/VCR cells by lipofectamine,respectively.Northern blot and Western blot were used to detect the expression of mRNA and protein in SGC7901/VCR and SGC7901 cells and transfectants, and drug sensitivity of transfectants for VCR, CDDP and 5-FU was analyzed with MTT assay.

Immuno-protective effect of tumor cell vaccine on Kunming mice bearing Ehrlich ascites tumor.

AIM:To evaluate the immunity of chemically modified tumor cell vaccine.METHODS:Tumor cell vaccines (TCV) were prepared by incubating the live Ehrlich ascites tumor cells with concanavalin A-mitomycin C (ConA-MMC), mitomycin C (MMC), concanavalin A-glutaraldehyde (ConA-Glu), glutaraldehyde (Glu), or paraformaldehyde (Para), respectively. The whole cell or soluble forms of the vaccines were administered intraperitoneally into Kunming mice once a week for three times prior to the intraperitoneal inoculation of a lethal dose of live tumor cells.

Two novel gastric cancer-associated genes identified by differential display.

AIM:To clone novel gastric cancer-associated genes and investigate their roles in gastric cancer occurrence.METHODS:A method called differential display was used which allows the identification of differentially expressed genes by using PAGE to display PCR-amplified cDNA fragments between gastric cancer cells and normal gastric mucosa cells. These fragments were cloned into plasmid vector pUC18.

IGF-1 induces Pin1 expression in promoting cell cycle S-phase entry.

Insulin-like growth factor I (IGF-1) is a well-established mitogen to many different cell types and is implicated in progression of a number of human cancers, notably breast cancer. The prolyl isomerase Pin1 plays an important role in cell cycle regulation through its specific interaction with proteins that are phosphorylated at Ser/Thr-Pro motifs. Pin1 knockout mice appear to have relatively normal development yet the Pin1(-/-)mouse embryo fibroblast (MEF) cells are defective in re-entering cell cycle in response to serum stimulation after G0 arrest.