Objective To study the function of vascular endothelial growth factor (VEGF) in type 2 diabetes model rats and its effect on focal cerebral ischemia induced by middle cerebral artery occlusion in these rats. Methods Focal cerebral ischemia was induced by middle cerebral artery occlusion for 6 hours in type 2 diabetes rats and normal control rats.Blood vessels morphology was examined by ink perfusion,infarct size was measured by TTC and expression of VEGF and CD34 were evaluated by immunohistochemistry staining. Results Ink perfusion revealed increased number of small vessels in type 2 diabetes rats. Infarct size was significantly smaller in type 2 diabetes rats ( ( 80. 07 ± 11.21 ) mm3 ) than that in normal controls ((98. 91 ± 14. 86) mm3,t = 2.48,P = 0. 0326). There were more hemorrhage lesions in the ischemic hemisphere in type 2 diabetes rats when comparing with the controls. VEGF and CD34 showed significantly higher expression in type 2 diabetes rats than in normal controls. Conclusions High expression of VEGF and CD34 are found in type 2 diabetes rats after middle cerebral artery occlusion. There is cerebrolvascular remodeling in diabetes rats. While this diabetes-induced remodeling appears to prevent infarct expansion,the changes also increase the risk of hemorrhagic transformation. The latter may result in poor prognosis.

OBJECTIVETo examine the temporal and spatial expression of vascular endothelial growth factor (VEGF) and angiopoietins (Ang) in rat brain after cerebral ischemia, and to elucidate the roles they played in angiogenesis and vascular permeability.

METHODSRats were subjected to either middle cerebral artery occlusion (MCAO) or sham operation. Reverse transcriptase-polymerase chain reaction, Western blotting, and immunohistochemistry were used to detect the expression of VEGF, Ang-1 and Ang-2 at different time points after ischemia. CD31 was used to label endothelial cells after MCAO. Vascular permeability was determined by Evans blue.

RESULTSVEGF was markedly increased at 2 h, had an initial peak at 12 h (0.7249 ± 0.1933, P < 0.01), and a second peak at 7 days (0.5264 ± 0.1519, P < 0.01). Ang-2 mRNA and protein significantly increased after MCAO, both of them peaked at 12 h (0.6747 ± 0.2416, P < 0.01; 1.1197 ± 0.1780, P < 0.01). In contrast, Ang-1 mRNA and protein gradually decreased after MCAO, respectively reaching a minimum at 3 d (0.3220 ± 0.1427, P < 0.01) and 1 d (0.1298 ± 0.0293, P < 0.01). Changes in the expression of these factors correlated with the progress of angiogenesis and vascular permeability. Evans blue test revealed that the vascular permeability gradually increased, and peaked at day 1 after ischemia [(6.219 ± 0.887) µg/g, P < 0.01].

CONCLUSIONDynamic temporal changes in VEGF, Ang-1 and Ang-2 expression stimulate the cerebral angiogenesis after focal cerebral ischemia.

Angiopoietin-1 ; genetics ; metabolism ; Angiopoietin-2 ; genetics ; metabolism ; Animals ; Blotting, Western ; Capillary Permeability ; Immunohistochemistry ; Infarction, Middle Cerebral Artery ; metabolism ; pathology ; physiopathology ; Male ; Neovascularization, Physiologic ; RNA, Messenger ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

BACKGROUND/AIMS: Rats with a spontaneous null mutation in endothelin receptor type B or Ednrb (sl/sl; spotting lethal) lack enteric neurons in the distal bowel and usually die within the first week after birth. This early postnatal lethality limits their use for examining the potential of cell therapy to treat Hirschsprung disease, and for studies of the influence of EDNRB on the mature CNS and vascular systems. METHODS: We have developed a surgical intervention to prolong the life of the spotting lethal sl/sl rat, in which we perform a colostomy on postnatal (P) day 4-6 rats to avoid the fatal obstruction caused by the lack of colonic enteric neurons. RESULTS: The stomas remained patent and functional and the rats matured normally following surgery. Weight gains were comparable between control and Hirschsprung phenotype (sl/sl) rats, which were followed until 4 weeks after surgery (5 weeks old). We confirmed the absence of enteric neurons in the distal colon of rats whose lives were saved by the surgical intervention. CONCLUSIONS: This study provides a novel approach for studying EDNRB signalling in multiple organ systems in mature rats, including an animal model to study the efficacy of cell therapy to treat Hirschsprung disease.
Animals ; Cell- and Tissue-Based Therapy ; Colon ; Colostomy ; Enteric Nervous System ; Female ; Hirschsprung Disease* ; Metrorrhagia ; Models, Animal* ; Neurons ; Parturition ; Phenotype ; Rats* ; Receptors, Endothelin ; Weight Gain

OBJECTIVETo investigate UbcH10 expression in hepatocellular carcinoma and explore its clinicopathological implications.

METHODSWe detected UbcH10 mRNA expression using RT-PCR in normal liver cell line, cancer cell lines, surgically removed hepatocellular carcinoma tissue and corresponding adjacent non-tumor tissue and evaluated the clinicopathological significance of UbcH10. Immunohistochemistry was performed to investigate UbcH10 protein expression in hepatocellular carcinoma tissue, the adjacent tissue, and normal liver tissue specimens.

RESULTSNormal liver cell line L02 showed significantly lower UbcH10 mRNA expression levels than the cancer cell lines BEL-7402, Hep3B, HepG2 and SMMC-7721 (P<0.05). UbcH10 mRNA expression was also was significantly higher in hepatocellular carcinoma tissues than in the corresponding non-tumor tissues (P<0.05). Clinicopathological evaluation suggested that UbcH10 expression was associated with tumor invasion of the portal vein, tumor size, TNM staging, and tumor differentiation (P<0.05). Immunohistochemistry identified stronger UbcH10 expression in hepatocellular carcinoma tissues than in the adjacent tissues and normal liver tissues (68.6%, 28.6%, and 26.7%, respectively).

CONCLUSIONUbcH10 is over-expressed in hepatocellular carcinoma and may serve as a novel biomarker as well as a therapeutic target of hepatocellular carcinoma.

Adult ; Aged ; Biomarkers, Tumor ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Female ; Hep G2 Cells ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Ubiquitin-Conjugating Enzymes ; genetics ; metabolism