Tumor-specific expression of shVEGF and suicide gene as a novel strategy for esophageal cancer therapy.

AIM: To develop a potent and safe gene therapy for esophageal cancer. METHODS: An expression vector carrying fusion suicide gene (yCDglyTK) and shRNA against vascular endothelial growth factor (VEGF) was constructed and delivered into EC9706 esophageal cancer cells by calcium phosphate nanoparticles (CPNP). To achieve tumor selectivity, expression of the fusion suicide gene was driven by a tumor-specific human telomerase reverse transcriptase (hTERT) promoter. The biologic properties and therapeutic efficiency of the vector, in the presence of prodrug 5-fluorocytosine (5-FC), were evaluated in vitro and in vivo. RESULTS: Both in vitro and in vivo testing showed that the expression vector was efficiently introduced by CPNP into tumor cells, leading to cellular expression of yCDglyTK and decreased VEGF level. With exposure to 5-FC, it exhibited strong anti-tumor effects against esophageal cancer. Combination of VEGF shRNA with the fusion suicide gene demonstrated strong anti-tumor activity. CONCLUSION: The shVEGF-hTERT-yCDglyTK/5-FC system provided a novel approach for esophageal cancer-targeted gene therapy.

Osteopontin expression is associated with hepatopathologic changes in Schistosoma japonicum infected mice.

AIM: To investigate osteopontin expression and its association with hepatopathologic changes in BALB/C mice infected with Schistosoma japonicum. METHODS: The schistosomal hepatopathologic mouse model was established by abdominal infection with schistosomal cercaria. Liver samples were obtained from mice sacrificed at 6, 8, 10, 14, and 18 wk after infection. Liver histopathological changes were observed with hematoxylin-eosin and Masson trichrome staining. The expression of osteopontin was determined with immunohistochemistry, reverse transcription-polymerase chain reaction, and Western blotting. The expression of α-smooth muscle actin (α-SMA) and transforming growth factor-ß1 (TGF-ß1) were determined by immunohistochemistry. Correlations of osteopontin expression with other variables (α-SMA, TGF-ß1, hepatopathologic features including granuloma formation and degree of liver fibrosis) were analyzed. RESULTS: Typical schistosomal hepatopathologic changes were induced in the animals. Dynamic changes in the expression of osteopontin were observed at week 6. The expression increased, peaked at week 10 (P < 0.01), and then gradually decreased. Positive correlations between osteopontin expression and α-SMA (r = 0.720, P < 0.01), TGF-ß1 (r = 0.905, P < 0.01), granuloma formation (r = 0.875, P < 0.01), and degree of liver fibrosis (r = 0.858, P < 0.01) were also observed. CONCLUSION: Osteopontin may play an important role in schistosomal hepatopathology and may promote granuloma formation and liver fibrosis through an unexplored mechanism.

[The diagnostic value of double-balloon enteroscopy in 67 cases with obscure abdominal pain].

OBJECTIVE: To evaluate the diagnostic valve of double balloon enteroscopy in patients with obscure abdominal pain and analyze the etiology of chronic abdominal pain resulted from enteral diseases. METHODS: Sixty-seven cases with chronic abdominal pain underwent a previous negative gastroscopy, colonoscopy, gastrointestinal barium, B ultrasound and electrocardiogram were received double balloon enteroscopy during June 2005 to June 2008. RESULTS: Thirty-six of 67 patients was done by enteroscopy via anus, and 19 cases via oral, and 12 cases via both anus and oral. The lesions were found in 41 of the 67 patients, with overall diagnostic yield of 61.19%. Among 41 cases of abdominal pain resulted from small bowel diseases, Crohn's disease were found in 15 cases (36.59%), non-specific small enteritis in 10 cases (24.39%), tumors in 8 cases (19.51%), other enteral diseases in 8 cases (19.51%). CONCLUSIONS: Double balloon enteroscopy was a diagnostic modality with a high diagnostic value for obscure abdominal pain resulted from small bowel diseases. The most common causes of obscure abdominal pain were Crohn's disease, non-specific small enteritis and tumors.

[Establishment of protein expression profile of human normal colonic epithelia].

OBJECTIVE: To establish a protein expression profile of human normal colonic epithelia. METHODS: Two-dimensional gel electrophoresis (2-DE) was applied to separate the total proteins of 20 human normal colonic epithelial tissues. The expression proteins in the human normal colonic epithelia were identified by both matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and electrospray ionization tandem mass spectrometry (ESI-Q-TOF), and the biological function and subcellular locations of the identified proteins were analyzed by bioinformatics. RESULTS: A 2-DE reference map of human normal colonic epithelium was established. On the 2-DE map, 1020+/-50 protein spots were detected, 204 protein spots representing 162 non-redundant proteins were identified, and 37 proteins had posttranslational modification. The identified proteins were categorized into several protein groups according to their functions or subcellular locations, whose data were available at our website (http://www.xyproteomics.org). CONCLUSION: A protein expression profile of human normal colonic epithelia is established for the first time, which provides useful information for investigating the physiological functions and pathologic process of colonic epithelia.

[Identification of aging related proteins in human normal colonic epithelium].

OBJECTIVE: To explore the molecular mechanisms of colonic epithelial aging related proteins and aged colonic epithelial susceptibility to tumor. METHODS: The proteins of normal human colonic epithelial tissue from young and old people were separated by 2-dimensional gel electrophoresis (2DGE), respectively. Then gels were stained by silver, scanned by imagescanner and analyzed with PDQuest software. The differentially expressed protein spots of colonic epithelium between the old and the young groups were identified by peptide mass fingerprint based on matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and database searching. RESULTS: Well-resolved and reproducible 2DGE maps of normal human colonic epithelium from the young and the old were acquired. Nineteen more than 2 fold differentially expressed protein spots were identified representing 17 different proteins by MALDI-TOF-MS. The functions of these proteins involve in metabolism, energy generation, transportation, antioxidation, translation and protein folding. CONCLUSION: Seventeen aging related proteins of human colonic epithelium identified indicate that injury of mitochondrial function and decline of antioxidant capability are important reasons for the aging of human colonic epithelium. These data provided useful clues for elucidating the mechanisms of colonic epithelial aging and aged colonic epithelial susceptibility to cancer.