Acute lung injury induced by H9N2 virus in mice / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>H9N2 avian influenza viruses (AIVs) have repeatedly caused infections in mammals even humans in many countries. The purpose of our study was to evaluate the acute lung injury (ALI) caused by H9N2 viral infection in mice.</p><p><b>METHODS</b>Six- to eight- week-old female SPF C57BL/6 mice were infected intranasally with 1 × 10(4) MID50 of A/HONG KONG/2108/2003 [H9N2 (HK)] virus. Clinical signs, pathological changes, virus titration in tissues of mice, arterial blood gas, and cytokines in bronchoalveolar lavage fluid (BALF) and serum were observed at different time points after AIV infection.</p><p><b>RESULTS</b>H9N2-AIV-infected mice exhibited severe respiratory syndrome, with a mortality rate of 50%. Lung histopathological changes in infected mice included diffuse pneumonia, alveolar damage, inflammatory cellular infiltration, interstitial and alveolar edema, and hemorrhage. In addition, H9N2 viral infection resulted in severe progressive hypoxemia, lymphopenia, and a significant increase in interleukin 1, interleukin 6, tumor necrosis factor, and interferon in BALF and serum.</p><p><b>CONCLUSIONS</b>The results suggest that H9N2 viral infection induces a typical ALI in mice that resembles the common features of ALI. Our data may facilitate the future studies of potential avian H9N2 disease in humans.</p>

Therapeutic effects of replication-competent adenovirus carrying human endostatin gene on pancreatic cancer in mice / 中华肝胆外科杂志

Objective To establish human pancreatic cancer xenografts in nude mice, and to investigate the antitumor efficacy of human endostatin expressed by replication-competent adenovirus AdTPHre-hE in vivo. Methods Pancreatic cancer cells AsPC-1 were injected subcutaneously in BALB/c nude mice to establish the xenografts. Tumor growth was observed and measured after AdTPHre-hE treatment. Expression of endostatin was detected by ELISA assay. The tumors were harvested for pathologic examination and immunohistochemical staining. Results Tumors grew more slowly in the AdTPHre-hE group and their sizes were markedly smaller than those of the Ad-hE group (P＜0.01)and control group(P＜0. 01). Endostatin levels were detected in the sera of nude mice in all treated groups, and endostatin expression in AdTPHre-hE group increased with time. The endostatin level in the AdTPHre-hE treated group was much higher(P＜0. 01)and increased faster than that in the Ad-hE treated group. Immunohistochemical staining for Hexon of adenovirus capsid showed more positive tumor cells in the tumor tissues treated with AdTPHre-hE. Immunohistochemical staining for FⅧ revealed a decreased microvessel density in the tumor tissues treated with AdTPHre-hE. Conclusion The replication-competent adenovirus efficiently expressed high-level endostatin and significantly inhibited tumor growth in vivo.

Construction and identification of a double-regulated replicative adenovirus AdTPHre-hEndo / 中华肝胆外科杂志

Objective To develop a double-regulated replicative adenovirus carrying the Human endostatin gene(hEndo). Methods The plasmid pTPHre-hEndo was constructed by gene engineering technique, carrying human endostatin gene, in which El A gene and E1B gene were driven by human telomerase reverse transcriptase (hTERT) promoter and hypoxia response element (HRE) promoter,respectively. The pTPHre-hEndo was co-transfected with pBHGE3 in 293 cells to generate recombinant adenovirus AdTPHre-hEndo. Virus titer was measured by the TCID50 method. Virus replication assay was performed to evaluate the selective replication ability of AdTPHre-hEndo. The transgene expression of endostatin was detected by ELISA assay. Results A novel gene-viral therapeutic system AdTPHre-hEndo was constructed by gene engineering technique and its titer was 3. 25 X 1010 pfu/ml.Proliferative test revealed that AdTPHre-hEndo could proliferate selectively in telomerase positive tumors. Furthermore, in comparison with non-replicative adenovirus Ad-hEndo, the transgene expression of endostatin mediated with AdTPHre-hEndo was significantly increased (P < 0. 01).Conclusion The novel gene-viral therapeutic system AdTPHre-hEndo has the capacity to replicate in pancreatic cancer cells and expresses the endostatin efficiently, and may provide a new strategy for pancreatic cancer gene therapy.

Treatment of pancreatic cancer by replicating adenovirus mediated human endostain gene in nude mice / 中华普通外科杂志

Objective To investigate the effect of a dual regulation of replicating adenovirus vector carrying human endostatin gene (AdTPHre-hEndo) on pancreatic cancer. Methods Human endostatin (hEndo) gene was cloned into the genome of replicating adenovirus specific for the tumor cells by virus recombination technology. The virus titer was 3.25 × 1010pfu/ml. A Balb/c nude mouse model carring sw1990 cells pancreatic cancer was established, the expression of human endostain and inhibition of tumor cells in vivo were detected. Results We successfully constructed AdTPHre-hEndo. The inhibition on pancreatic cancer cell line SW-1990 of AdTPHre-hEndo is better than Ad-hEndo (P ＜0. 01 ), and ONYX-015 ( P ＜ 0. 05 ). The endostatin expression of AdTPHre-hEndo group was significantly higher than Ad-hEndo group and the control group (P ＜ 0. 01 ). The intratumoral MVD also decreased significantly in the treated tumors(6. 8 ±2. 5 vs. 16. 0 ±4. 6、47. 2 ± 10. 0, P ＜0. 01 ). Conclusions The recombination adenovirus can express biologically active hEndo effectively, which results in inhibiting the growth of micro-blood vessels and proliferation slowly.

Effect of wild type parkin gene on growth of hepatocellular carcinoma cells / 中华肝胆外科杂志

Objective To explore the effect of wild type or mutant parkin gene expression on the growth of human hepatocellular carcinoma cell line Huh-7. Methods The parkin (wild type or mutant) expression vector and empty vector were transferred into Huh-7 cell lines with LipofectAMINE 2000 reagents. The positive clones that expressed parkin gene stably were chosen by G418 and checked by reverse transcription-polymerase chain reaction (RT-PCR) to check the DNA sequences. The cytobiological behaviors of those positive clones were analyzed by cell proliferation assay and tumorigenesis in nude mice. Results Huh-7 cell lines that expressed wild type or mutant parkin gene stably were successfully established. The growth of wild type parkin-expressed cells was obviously inhibited compared with the control cells transfected with empty vectors(t= 3. 875, P= 0. 031).The volume of tumor formed by wild type parkin-expressing cells in nude mice was also significantly reduced (t=8. 228,P=-0. 003). Mutant parkin gene expression had a slight effect on the growth of Huh-7 cells in vitro and in vivo (P＞0.05). Conclusion The re-expression of wild type parkin gene can favor the malignant phenotype revision of Huh-7 cells. Therefore, it might be a good candidate for tumor suppressor gene associated with HCC.

PI_3 K/Akt signal effects Ephrin-A1 mediated malignancy of HCC cells / 中华普通外科杂志

Objective To investigate the role of PI_3 K/Akt signal pathway in Ephrin-Al gene mediated invasion,metastasis of Huh-7 cells.Methods Western blot was used to test the protein expression of phosphatidylinositol 3-kinase(PI_3 K)and mitogen-activated protein kinase(MAPK)after Huh-7 cells were treated with Ephrin-A1/Fc fusion protein.According to the protein expression,LY294002 was used to block PI_3 K/Akt pathway specifically,then p-Akt protein expression,mobility and invasive ability of Huh-7 cells were examined.Results In Huh-7 cells actived by Ephrin-Al/Fc fusion protein,p-Akt expression was higher than that in control group(t=4.564,P＜0.05),but there was no difference of p-p38MAPK expression between Ephrin-Al/Fc fusion protein group and IgG/Fc fusion protein group(P＞0.05).PI_3 K/Akt pathway was specifically blocked by LY294002,the p-Akt protein expression decreased in Huh-7 cells,and the mobility and invasive ability mediated by Ephrin-Al in Huh-7 cells decreased(P＜0.05).Conclusions PI_3 K/Akt pathway effects an important role in mobility and invasive ability of Huh-7 cells mediated by Ephrin-A1.

Mutations analysis in a pedigree with maternally inherited sensorineural hearing loss / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze the mutations in a pedigree with maternally inherited sensorineural hearing loss, and to investigate whether 235delC heterozygote mutation in gap junction protein beta 2 (GJB2) gene modulates the severity of hearing loss associated with the A1555G mitochondrial mutation.</p><p><b>METHODS</b>The PCR products were digested with the Alw26 I restriction enzyme, followed by direct sequencing to detect the mitochondrial mutations in 72 members of a core pedigree of an extensive family with matrilineal nonsyndromic deafness; 235delC mutation of the GJB2 gene was screened in this family by using the Apa I restriction enzyme and direct sequencing.</p><p><b>RESULTS</b>The A1555G mutation of the mitochondrial DNA was present in all 27 members of maternal line, out of them, 21 members had phenotype of deafness (77.8%), with a high penetrance. Only three maternal line members of 72 members possessed 235delC heterozygote mutations, and the three had different phenotypes.</p><p><b>CONCLUSION</b>The A1555G homozygous mutation of mitochondrial DNA is the susceptive etiological factor of nonsyndromic deafness in this family, but in the study of this pedigree, the 235delC heterozygous mutation in GJB2 gene may not aggravate the symptoms of hearing loss associated with the A1555G mitochondrial mutation.</p>

Adenoviral vector encoding the whole humanized antibody gene of preS_2 antigen inhibiting the recurrence of hepatitis B virus after liver transplantation / 中华器官移植杂志

Objective To investigate the adenoviral vector encoding the whole humanized antibody gene of pres_2 antigen for preventing the liver graft from infection by the hepatitis B virus. Methods The whole humanized antibody gene to pres_2 antigen was cloned into type 5 adenoviral shuttle plasmid pDC315. The corresponding recombinant virus was obtained by homologous recombination in 293 packaging cells. The virus containing the whole humanized antibody gene of preS_2 antigen was transfected into the rat liver graft during cold preservation. The effect of the recipient serum containing preS_2 antibody protecting human hepatocytes from hepatitis B virus infection was observed. Results The viral titer determined by TCID50 analysis was 5.1 ?10 10 PFU/ml. The concentration of preS_2 antibody was ( 16.7 ? 10.5 ) mg/L on the day 3 and ( 30.9 ? 13.6 ) mg/L on the day 7. The serum containing preS_2 antibody could protect human hepatocytes from hepatitis B virus infection in vitro when the concentration of the preS_2 antibody was more than 0.5 ?g/ml. Conclusions The adenoviral vector encoding the whole humanized antibody gene of preS_2 protein were constructed successfully. The whole humanized antibody was expressed. The preS_2 antibody in recipient serum protecting human hepatocytes from hepatitis B virus infection was observed, which might be a new method to prevent hepatitis B re-infection after liver transplantation.