Aquaporin-9 downregulation prevents steatosis in oleic acid-induced non-alcoholic fatty liver disease cell models.

Aquaporin-9 (AQP9) is an aquaglyceroporin that acts as the adipose glycerol channel. However, the role of AQP9 in steatosis in non-alcoholic fatty liver disease (NAFLD) has not yet been fully elucidated. In the present study, the coding sequence of the AQP9 gene was obtained from LO2 cells by RT-PCR, and cloned into the pEGFP-N1 vector.

[Construction of short hairpin RNA targeting aquaglyceroporin 9 and screening its effect on molecular mechanisms of nonalcoholic fatty liver disease using a cell model system].

Objective: To construct a short hairpin (sh)RNA targeting aquaglyceroporin 9 (AQP9) that effectively silences gene expression in liver cells in order to investigate of the role of AQP9 in nonalcoholic fatty liver disease (NAFLD) pathogenesis using an in vitro cell model system.

Methods: Small interfering (si)RNAs were designed against the human gene sequences encoding AQP9 (NCBI GenBank Accession No. AB008775) and unrelated control sequences, synthesized, annealed to form double-strands, and inserted into the pGenesil- 1 shRNA-expression plasmid.

Oral immunization with recombinant Mycobacterium smegmatis expressing the outer membrane protein 26-kilodalton antigen confers prophylactic protection against Helicobacter pylori infection.

Helicobacter pylori infection is prevalent worldwide and results in chronic gastritis, which may lead to gastric mucosa-associated lymphoid tissue lymphoma and gastric cancer. We have previously reported that oral immunization with recombinant Mycobacterium smegmatis expressing the H. pylori outer membrane protein 26-kilodalton (Omp26) antigen affords therapeutic protection against H.

Attenuated Salmonella typhimurium carrying TRAIL and VP3 genes inhibits the growth of gastric cancer cells in vitro and in vivo.

Background: Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand (TRAIL) and apoptin (VP3) of chicken anemia virus can selectively induce apoptosis in human tumor cell lines by two different pathways. Salmonella not only delivers functional genes to mammalian cells but also possesses antitumor activity and therefore could be adopted as a novel vector for anticancer therapy.

Materials And Methods: TRAIL and VP3 genes were cloned into a pBudCE4.

[Overexpression of adiponectin prevents hepatocyte steatosis].

Objective: To investigate the effect of adiponectin on hepatocyte steatosis.

Methods: L02 cells were transfected with pEGFP-N1-AdipoQ, a plasmid encoding pEGFP-adiponectin fusion protein, or pEGFP-N1. Lipid droplets in the hepatocytes were observed by oil red staining at 72 h.

[Anti-tumor effect of eukaryotic expressing plasmid containing soluble tumor necrotic factor-related apoptosis inducing ligand combined with human angiostatin Kringle (1 - 3) genes on human gastric cancer xenografts in nude mice].

Objective: To investigate the anti-tumor effect of eukaryotic expressing plasmid containing human angiostatin Kringle (1 - 3) [hAG (K1-3)] combined with soluble tumor necrotic factor-related apoptosis inducing ligand (sTRAIL) genes on human gastric cancer xenografts in nude mice.

Methods: Recombinant plasmids of pBud-hAG and pBud-hAG-TRAIL were constructed by subcloning technique. Twenty nude BALB/c mice were inoculated with human gastric cancer cells of the line BGC-823 subcutaneously into the back.

Recombinant Mycobacterium smegmatis mc(2)155 vaccine expressing outer membrane protein 26 kDa antigen affords therapeutic protection against Helicobacter pylori infection.

Orally administered recombinant Mycobacterium smegmatis (rM. smegmatis) vaccines represent an attractive option for mass vaccination programmes against various infectious diseases. Therefore, in the present study, we evaluated the capacity of the outer membrane protein 26kDa antigen (Omp26) of Helicobacter pylori (H.

RNA interference-mediated silencing of the Hsp70 gene inhibits human gastric cancer cell growth and induces apoptosis in vitro and in vivo.

Aims And Background: The role of heat shock protein (HSP) 70 in gastric cancer has been extensively examined in many studies for the past decade. It has been demonstrated that over-expression of Hsp70 might play important role in malignant transformation and maintenance of malignant phenotypes. Therefore, silencing the Hsp70 gene could be applicable in molecular therapies of human gastric cancer.

Comparison of esomeprazole enteric-coated capsules vs esomeprazole magnesium in the treatment of active duodenal ulcer: a randomized, double-blind, controlled study.

Aim: To evaluate the efficacy and tolerability of two different preparations of esomeprazole in healing duodenal ulcers.

Methods: A total of 60 patients with active duodenal ulcers were enrolled and randomized to receive esomeprazole enteric-coated capsules (40 mg) or esomeprazole magnesium (40 mg), once daily, for 4 consecutive wk, with ulcer healing being monitored by endoscopy. Safety and tolerability were also assessed.

[Effects of pshRNA-DNMT1 on the proliferation and apoptosis of gastric cancer: experiments in vitro and in vivo].

Objective: To investigate the effects of pshRNA-DNMT1 on the proliferation and apoptosis of gastric cancer.

Methods: Recombinant eukaryotic expression plasmid pshRNA-DNMT1 containing the sequence of the gene of DMNT1 that methylates the specific pyrimidine residue in the DNA promoter region was constructed. Human gastric cells of the line AGS were cultured and transfected with pshRNA-DNMT1.

[Effects of dnmt1 gene silencing on cell cycle, proliferation, and apoptosis of gastric cancer cell line AGS].

Background & Objective: Dnmt1, a major DNA methyltransferase gene, is highly expressed in many cancers and lowly expressed in normal adult cells, therefore, its overexpression is closely related to tumorigenesis. This study was to assess effects of dnmt1 gene silencing on cell cycle, proliferation, and apoptosis of gastric cancer cell line AGS.

Methods: The eukaryotic expression plasmid pshRNA-dnmt1, containing the sequence of short hairpin RNA (shRNA) targeting dnmt1, was constructed and transfected into AGS cells.

[Establishment of cellular hypoxia model and its effects on biological behavior of gastric cancer cell line SGC-7901].

Background & Objective: Hypoxia is an elementary characteristic of tumor microenvironment. Establishing hypoxia microenvironment in vitro will be of help in studying the effects of hypoxia on tumor cells. Previous methods of establishing hypoxic cell culture model are fussy, and researches on the effects of hypoxia on gastric cancer cells are rare.

Diagnosis of Helicobacter pylori infection and diseases associated with Helicobacter pylori by Helicobacter pylori outer membrane proteins.

Aim: To examine the serological response of patients with upper gastrointestinal diseases and Helicobocter pylori (H pylori) infection to two H pylori outer membrane proteins (OMPs) (Mr18,000 and Mr 26,000) acquired by gene recombinant technique, and to determine the diagnostic significance of serological tests derived from these OMPs.

Methods: Recombinant vectors encoding the two H pylori OMPs were used to transform and express in BL21 (DE3) E.coli.

Cloning and sequence analysis of gene oipA encoding an outer membrane protein of human Helicobacter pylori.

Aim: To construct a recombinant E. coli strain that would highly express the proinflammatory outer membrane protein of human Helicobacter pylori (H pylori).

Methods: The oipA DNA was amplified by PCR, inserted into pET-32a, and transformed into Top10 E.

[Construction, expression and antigenicity of bivalent vaccine candidate of human Helicobacter pylori].

Aim: To construct a recombinant vector containing fused gene of heat shock protein A(HspA) and outer membrane protein(OMP) with M(r) 18,000, from human Helicobacter pylori(Hp) and express the fusion protein in E.coli BL21.

Methods: The gene encoding HspA was amplified from Hp chromosome by PCR.

Construction and characterization of bivalent vaccine candidate expressing HspA and M(r)18,000 OMP from Helicobacter pylori.

Aim: To construct a recombinant vector which can express outer membrane protein (OMP) with M(r)18,000 and heat shock protein A (HspA) from Helicobacter pylori (H. pylori) in E. coli BL21, and to exploit the possibility for obtaining the vaccine conferring protection from H.

[Construction, expression and antigenic study of bivalent vaccine candidate with 26,000 OMP and heat short protein A of human Helicobacter pylori].

Objective: To construct a recombinant vector containing gene encoding heat shock protein A (HspA) and outer membrane protein (OMP) with relative molecule mass (Mr) of 13,000 and 26,000 respectively from human Helicobacter pylori (Hp) and be expressed in E. coli BL21, as well as analyse its antigenic for the exploiting vaccine of Hp.

Methods: The target gene encoding heat shock protein A was amplified from Hp chromosome by PCR.

Effect of compound rhodiola sachalinensis A Bor on CCl4-induced liver fibrosis in rats and its probable molecular mechanisms.

Aim: To explore the anti-fibrotic effect of a traditional Chinese medicine, compound rhodiola sachalinensis A Bor on CCl(4)-induced liver fibrosis in rats and its probable molecular mechanisms.

Methods: Ninety healthy male SD rats were randomly divided into three groups: normal group (n=10), treatment group of compound rhodiola sachalinensis A Bor (n=40) and CCl(4)-induced model group (n=40). The liver fibrosis was induced by CCl(4) subcutaneous injection.

A study of recombinant protective H.pylori antigens.

Aim: To construct a recombinant vector which can express M (r)26000 outer membrane protein (OMP) from Helicobacter pylori (Hp), and to obtain the vaccine protecting against Hp infection and a diagnostic reagent kit quickly detecting Hp infection.

Methods: The gene encoding the structural M(r)26000 outer membrane protein of Hp was amplified from Hp chromosomal DNA by PCR, and inserted in the prokaryotic expression vector pET32a (+), which was transformed into the Top10 E. coli strain.