Autophagy is involved in recombinant Newcastle disease virus (rL-RVG)-induced cell death of stomach adenocarcinoma cells in vitro.

Oncolytic viruses can kill malignant cells while sparing normal cells. Multiple pathways are involved in this action. The antitumor effects of viral infection on SGC-7901 and AGS cells were investigated. We measured endoplasmic reticulum stress and autophagy caused by the recombinant avirulent Newcastle disease virus (NDV) LaSota strain expressing the rabies virus glycoprotein (rL-RVG) and the NDV wild-type strain. The dose-response curves were analyzed using the MTT assay. The expression of RVG was detected by western blotting, RT-PCR and immunofluorescence analyses. Cell death and autophagy were observed using transmission electron microscopy, TUNEL and western blotting. Endoplasmic reticulum stress and the mitochondrial transmembrane potential were detected by western blotting and immunofluorescence, respectively. Immunofluorescence, western blot and RT-PCR analyses indicated that RVG gene and protein were expressed in SGC-7901 and AGS cells infected by rL-RVG. MTT and TUNEL analyses showed that the growth of SGC-7901 and AGS cells in the rL-RVG-infected group was significantly inhibited compared with the wild-type NDV-infected group (p<0.05). Western blot analysis indicated that rL-RVG and NDV induced increases in apoptosis, endoplasmic reticulum stress, and autophagy in the SGC-7901 and AGS cells. However, apoptosis and autophagy decreased in these cells after the application of the autophagy pathway inhibitor 3-MA or ATG-5-specific siRNA. Immunofluorescence analysis showed that the mitochondrial membrane potential collapsed. Taken together, these results indicate that the rL-RVG virus group is much more powerful compared with the NDV-infected group (p<0.05). rL-RVG and NDV are potent antitumor agents that induce autophagy.

[Pathology of myocardial fatty infiltration: an autopsy report from 117 cases].

OBJECTIVE: To explore the risk factors related to the formation of myocardial fatty infiltration and possible pathological consequences. METHODS: The macroscopic and microscopic findings in 117 autopsy cases with myocardial fatty infiltration were examined during October, 2001 to June, 2009. RESULTS: There was a significant positive correlation between the macroscopic grading of subepicardial adipose tissue and the microscopic myocardial fatty infiltrative degree(r(s) = 0.57, P < 0.01) but there was no correlations between the myocardial fatty infiltrative degree and age as well as coronary arteriosclerosis (all P > 0.05). The percent of myocardial atrophy was 39.32% (46/117), and the rate of myocardial atrophy in mild myocardial fatty infiltration group (13/63, 20.63%) was significantly lower than that in moderate myocardial fatty infiltration group (22/39, 34.92%; chi(2) = 12.14, P < 0.01) and in severe myocardial fatty infiltration group (11/15, 73.33%; chi(2) = 13.42, P < 0.01). There were 28 sudden cardiac deaths among the 117 cases including 6 deaths due to myocardial fatty infiltration. CONCLUSIONS: Myocardial fatty infiltration is often associated with myocardial atrophy, even with sudden cardiac death but is not an accompanying pathologic changes of aging and coronary arteriosclerosis.

[Pathology of accidental electrocution: an autopsy study of 16 cases].

OBJECTIVE: To study the pathologic findings seen in lethal cases due to accidental electrocution. METHODS: The macroscopic and microscopic findings in 16 autopsy cases died of electrocution encountered during the period from January, 2001 to July, 2008 were retrospectively reviewed. RESULTS: Typical electric marks were found on gross examination in 5 of the 16 cases studied. Histologically, 11 of the 16 cases showed evidence of electric burn. The morphologic features of atypical electric marks varied. Simple epidermal exfoliation and color changes were relatively common. Pathologic changes in internal viscera included disarray of myocardial fibers. Rupture of myocardial fibers was readily identified than in non-electrocution death. Sometimes, focal interstitial hemorrhage and polarization of endothelial cells were seen. CONCLUSIONS: The electric marks on the skin, as confirmed by histologic examination, remain important sequelae of electrocution. The pathologic changes seen in myocardium provide additional clues to the diagnosis.

[Histopathological characterization of drug-related death].

OBJECTIVE: To study the histopathological changes in drug-related death cases in order to provide valuable information for its diagnosis. METHODS: Thirty cases of drug-related death were collected for systemic autopsy and histopathology examination. Ante mortem history and other informations of each case were also reviewed and analyzed. RESULTS: Injection marks, emaciation, asphyxia and histopathological changes in critical organs and tissues correlated with addiction behavior. In the 30 cases, 20% died of diseases, 33.3% acute drug intoxication, 26.7% quitting drug, 10% sudden death, and 10% outside violence. CONCLUSION: Systemic autopsy and histopathology examination in drug-related death are useful for determination of the cause of death in these cases.

[Construction and screening of a ribozyme targeting telomerase RNA and effects of telomerase ribozyme on proliferation and apoptosis of CNE-2Z cells].

BACKGROUND & OBJECTIVE: It was proved that telomerase is an important determinant in tumor progression and cell immortalization. Ribozyme is a special kind of trans-acting RNA with endonuclease activity and sequence-specific catalytic RNA molecules, which can cleave target RNA. It was reported that telomerase activity is present in human poorly-differentiated nasopharyngeal carcinoma (NPC) CNE-2Z cells. This study was designed to construct eukaryotic expression plasmids containing telomerase ribozyme (teloRZ)gene targeting the template region of human telomerase RNA (hTR) and then to transfect the plasmids into CNE-2Z cells by electroporation to investigate the effect of teloRZ on proliferation and apoptosis of those transfected CNE-2Z cells. METHODS: Hammer ribozyme gene teloRZ directed against telomerase RNA templet was designed and synthesized to serve as a telomerase inhibitor. Three different eukaryotic expression plasmids carried with the green fluorescent protein (GFP) reporter gene and puromycin-resistance gene and containing teloRZ gene were constructed. They were referred to as pGFPuro-teloRZ2.1, pGFPuro-teloRZ7.1, and pGFPuro-teloRZ7.7 and differed in the relative orientation of the genes for telomerase-ribozyme and puromycin-resistance. The CNE-2Z cells were transfected with three expression plasmids and control plasmid pPAT-GFP by electroporation. The expression of GFP was detected by fluorescent microscope; cellular proliferation index (PI) and apoptosis were investigated by flow cytometry analysis and fluorescence staining. RESULTS: PI of CNE-2ZGTR7.1 cells transfected by plasmid pGFPuro-teloRZ7.1 (25.100%+/-0.141%)was significantly lower than those of CNE-2Z cells untransfected by any plasmid (53.663%+/-16.981%),CNE-2ZG cells transfected by control plasmid pPAT-GFP (61.575%+/-5.166%),CNE-2ZGTR2.1 cells transfected by plasmid pGFPuro-teloRZ2.1 (61.500%+/-20.082%), and CNE-2ZGTR7.7 cells transfected by plasmid pGFPuro-teloRZ7.7 (59.400%+/-13.933%) (P< 0.01). GFP was detected in CNE-2ZG cells,CNE-2ZGTR7.1 cells, and CNE-2ZGTR7.7 cells;while there was no GFP expression in CNE-2Z cells and CNE-2ZGTR2.1 cells. The plasmid pGFPuro-teloRZ7.1 was selected from 3 plasmids for further experiments. Apoptosis could be observed in CNE-2ZGTR7.1 cells after 12 generations. There was no apoptosis occurring in CNE-2Z and CNE-2ZG cells. CONCLUSION: The teloRZ7.1 gene was electroporated successfully into CNE-2Z cells. TeloRZ7.1 can inhibit the proliferation and induce apoptosis of CNE-2Z cells. These findings suggest the potential application of ribozyme teloRZ7.1 as telomerase inhibitor.