A novel oncolytic adenovirus inhibits hepatocellular carcinoma growth / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

OBJECTIVE@#To evaluate the inhibitory role of a novel oncolytic adenovirus (OA), GP73-SphK1sR-Ad5, on the growth of hepatocellular carcinoma (HCC).@*METHODS@#GP73-SphK1sR-Ad5 was constructed by integrating Golgi protein 73 (GP73) promoter and sphingosine kinase 1 (SphK1)-short hairpin RNA (shRNA) into adenovirus serotype 5 (Ad5), and transfecting into HCC Huh7 cells and normal human liver HL-7702 cells. The expression of SphK1 and adenovirus early region 1 (E1A) was detected by quantitative real-time PCR (qRT-PCR) and western blot, respectively. Cell viability was detected by methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay, and apoptotic rate was determined by flow cytometry. An Huh7 xenograft model was established in mice injected intratumorally with GP73-SphK1sR-Ad5. Twenty days after injection, the tumor volume and weight, and the survival time of the mice were recorded. The histopathological changes in tumor tissues were observed by hematoxylin-eosin (HE) staining and transmission electron microscopy (TEM).@*RESULTS@#Transfection of GP73-SphK1sR-Ad5 significantly upregulated E1A and downregulated SphK1 in Huh7 cells, but not in HL7702 cells. GP73-SphK1sR-Ad5 transfection significantly decreased the viability and increased the apoptotic rate of Huh7 cells, but had no effect on HL7702 cells. Intratumoral injection of GP73-SphK1sR-Ad5 into the Huh7 xenograft mouse model significantly decreased tumor volume and weight, and prolonged survival time. It also significantly decreased the tumor infiltration area and blood vessel density, and increased the percentages of cells with nucleus deformation and cells with condensed chromatin in tumor tissues.@*CONCLUSIONS@#GP73-SphK1sR-Ad5 serves as a novel OA and can inhibit HCC progression with high specificity and efficacy.

The expression of connective tissue growth factor in mast cells in the development of pulmonary fibrosis / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate whether connective tissue growth factor (CGGF) is expressed in mast cells (MCs) in lung in the development of bleomycin (BLM)-induced pulmonary fibrosis.</p><p><b>METHODS</b>Thirty-two male SD rats were randomly divided into 2 groups: BLM group and control group (n=16). The rats in BLM group were received single intratracheal instillation of BLM (5 mg/kg), and the rats in control group received equal volume of 0.9% normal saline(NS) to BLM. The rats in each group were sacrificed for lung tissue sampling on day 14 and day 28 after intratracheal instillation respectively. As the index of the severity of pulmonary fibrosis, the content of hydroxyproline in lungs was analyzed by chloramine T method. Mast cells and CTGF expression in lungs were examined by toluidine blue stain and immunohistochemical assay respectively.</p><p><b>RESULTS</b>(1) On day 28 after intratracheal instillation of BLM, the content of hydroxyproline in lungs of rats was higher than that of control rats (P < 0.01). (2) Compared to control rats, the rats on day 14 and day 28 after instillation of BLM showed increased number of mast cells (Both P < 0.01) and up-regulated CTGF expression (Both P < 0.01). (3) No CTGF immuno-positive MCs were seen in the lungs of control rats whereas CTGF immuno-positive MCs were observed in the pathological areas in lungs of rats on day 14 and day 28 after BLM.</p><p><b>CONCLUSION</b>CTGF is expressed in MCs in lungs in the development of pulmonary fibrosis, which might be one of the mechanisms underling promoting effect of MCs on fibrosis in lung.</p>

Baicalin prevents the up-regulation of connective tissue growth factor in fibrotic lungs of rats / 生理学报

To clarify the mechanism underlying the preventive effect of baicalin (Bai) on fibrosis in lung, we investigated the influence of Bai on the up-regulation of connective tissue growth factor (CTGF) in fibrotic lungs. Male Sprague-Dawley (SD) rats were divided into four groups randomly: normal saline (NS)+NS group (a single intratracheal instillation of NS plus i.p. injection of NS), NS+Bai group (intratracheal instillation of NS plus i.p. injection of Bai), bleomycin (BLM)+NS group (intratracheal instillation of BLM plus i.p. injection of NS) and BLM+Bai group (intratracheal instillation of BLM plus i.p. injection of Bai). All the i.p. injections were performed once daily. On day 28 after intratracheal instillation of BLM or NS, the rats were sacrificed for lung tissue sampling. As the index of the severity of pulmonary fibrosis, the content of hydroxyproline in lungs was analyzed by chloramine T method. The expression levels of CTGF mRNA and protein in the lungs were detected by RT-PCR and immunohistochemistry, respectively. The results showed that, compared to the rats in NS+NS group, the rats in BLM+NS group showed increased hydroxyproline content and higher levels of CTGF mRNA and protein expressions (P<0.01), suggesting that BLM had induced fibrosis in lung and up-regulated CTGF expression in the fibrotic lungs. Administration of different dosages of Bai (6, 12.5 and 50 mg/kg per d, for 28 days) into the BLM-treated rats reduced the increased content of hydroxyproline, and ameliorated the up-regulation of CTGF mRNA and protein levels, respectively. These results suggest that Bai could prevent the up-regulation of CTGF expression in fibrotic lungs of rats receiving BLM instillation, which might be one of the mechanisms underlying the preventive effect of Bai on pulmonary fibrosis.

Intracerebroventricular administration of adenosine A1 receptor antisense oligodeoxynucleotide inhibites the neuroprotective effect of the cerebral ischemic preconditioning in rats / 中国应用生理学杂志

<p><b>AIM</b>To further explore the role of adenosine A1 receptor in the neuroprotective effect of cerebral ischemic preconditioning, the present study was undertaken to observe the effect of inhibiting expression of adenosine Al receptor with adenosine A1 receptor antisense oligodeoxynucleotide (ARA1 As-ODN) on the neuroprotective effect of cerebral ischemic preconditioning against delayed neuronal death (DND) normally induced by lethal brain ischemia.</p><p><b>METHOD</b>The rat 4-vessel occlusion global cerebral ischemic model was used. Forty-eight male Wistar rats with permanent occlusion of the bilateral vertebral arteries were divided into 8 groups: Sham, CIP, brain ischemic insult, CIP + brain ischemic insult, Distilled water + CIP + brain ischemic insult, ARA1 As-ODN, ARA1 As-ODN +CIP, ARA1 As-ODN+ CIP + brain ischemic insult(two doses of 10 nmol/5 microl and 20 nmol/5 microl were used) groups. ARA1 As-ODN was dissolved in distilled water and injected into the right lateral cerebral ventricle. To illustrate the profile of DND, histological grade (HG) and neuronal density (ND) in the CA1 region of the hippocampus were examined 7 d after the sham operation or the last time of ischemia under thionin staining.</p><p><b>RESULTS</b>The HG and ND in CIP group were similar to those in sham group. Brain ischemic insult induced obvious DND as represented with the increase in HG and decrease in ND significantly (P < 0.05 vs. sham and CIP groups). In CIP + ischemic insult group,no obvious DND was observed,which indicated that CIP protected pyramidal neurons against the ischemic insult.While the administration of ARA1 As-ODN in ARA1 As-ODN + CIP + brain ischemic insult group caused obvious increase in HG and decrease in ND compared with CIP + brain ischemic insult group (P < 0.05) in a dose dependent manner,which indicated that the neuroprotective effect of CIP against DND of hippocampal pyramidal neurons normally induced by ischemic insult was inhibited by the administration of ARA1 As-ODN.</p><p><b>CONCLUSION</b>The results further demonstrate the association of up-regulation of adenosine A1 receptors with the induction of CIP-mediated BIT.</p>