Objective To explore the activation of PI3k/Akt pathway of serum deprived IEC-6 cells by the serum of rats exposed to single radiation,burn or combined injury.Methods The IEC-6 cells were cultured in serum deprived media for 24 h,and stimulated by the serum of rats exposed to single radiation(~(60)Co ? ray at dose of 9 Gy),single burn(exposure to 5 kW tungsten-halogen light till whole body Ⅲ degree burn) or combined injury(burn first and radiation),and the cells stimulated by the serum from the normal rats and serum starved cells served as the control group.The total proteins of different group cells were extracted and the levels of phosphorylation of Akt were tested by Western blotting.The differentially expressed low mass proteins in the serums were detected by SELDI proteinchip technology,and primarily analyzed by related software as well as bioinformatic methods.Results The level of phosphorylation of Akt in the IEC-6 cells stimulated by serum from rats exposed to single radiation,single burn or combined injury was higher than in the cells stimulated by the serum from normal rats,in which the burn serum caused the highest level.As compared to burn rat serum,the serum of radiation and combined injury had 13 and 6 differentially expressed protein peaks respectively.Conclusion All the serums from rats exposed to different kinds of damage agents could activate the PI3K/Akt pathway of IEC-6 cells efficiently.The special components of burnt rat serum may contribute to the highest effect on the phosphorylation of Akt.

Objective To detect whether mice bone marrow mesenchymal stem cells(BMSCs)can contribute to the regeneration of hepatocytes in bone marrow chimeric mice.Methods Female recipient mice(C57BL/6J)underwent whole body gamma-ray irradiation with a dose of 10 Gy to ablate their bone marrow,followed by immediate tail vein injection of BMSCs isolated from male GFP transgenic mice.Animals were killed at different phase points:1 week,1 month,and 3 months.Using fluorescence microscope we directly observed GFP-positive cells in the liver frozen sections,and we also prepared the parafilm sections to detect the GFP-positive cells and the coexpression of GFP and Alb,CK18 by immunohistochemistry and immunofluorescence respectively.Results We found numerous GFP-positive cells in recipient mice liver at 1 week after BMSCs transplantation,some at 1 month and seldom at 3 months.There were some cells coexpressing GFP and Alb,CK18 at all the phase points.Conclusion Allotype BMSCs can differentiate into Alb and CK18 positive hepatocytes in bone marrow chimeric mice,which will become an ideal cell resource for liver tissue project.

Objective: To explore clinical efficacy and safety of Wumeiwan combined with triple therapy for treatment of chronic atrophic gastritis (CAG). Methods: According to the random indicator method, 113 patients with CAG were divided into control group (n=56) and treatment group (n=57). Patients of control group were treated with triple therapy, while treatment groupwere treated Wumeiwan combined with triple therapy. The two groups were treated for 3 months. Clinical effect was evaluated after treatment. The helicobacter pylori (Hp) conversion to negative of the two groups was compared and recorded. The Hp overcast conditions of the two groups were compared. The serum TNF-α, IL-6, IL-8 and peripheral blood CD3⁺, CD4⁺, CD8⁺, CD4⁺/CD8⁺ of the two groups before and after treatment were compared. The adverse reactions of the two groups during the treatment were compared. Results: Total effective rate of treatment group was 94.7% (54/57), which was significantly higher than the control group 82.14% (46/56), and the difference was statistically significant (χ²=4.401, P=0.036). Hp overcast rate of treatment group was 88.2% (45/51), which was significantly higher than the control group 48.0%(24/50), and the difference was statistically significant (χ²=18.883, P=0.000). After treatment, the serum TNF-α (1.43 ± 0.17 mg/L vs. 1.97 ± 0.22 mg/L, t=14.615), IL-6 (30.79 ± 3.65 ng/L vs. 41.13 ± 4.10 ng/L, t=14.166), IL-8 (7.52 ± 1.32 ng/L vs. 9.60 ± 1.77 ng/L, t=7.090) in the treatment group were lower than those in the control group (P<0.05). After treatment, the peripheral blood CD3⁺ (75.12% ± 16.44% vs. 67.33% ± 14.37%, t=2.680), CD4⁺ (39.02% ± 11.41% vs. 33.49% ± 10.61%, t=2.667), CD4⁺/CD8⁺ (1.58 ± 0.35 vs. 1.19 ± 0.32, t=6.179) in the treatment group were higher than those in the control group (P<0.05), the peripheral blood CD8⁺ (24.75% ± 9.69% vs. 28.12% ± 11.29%, t=1.704) in the treatment group were higher than those in the control group (P<0.05). There was no significantly difference of the adverse reaction rates of the two groups during treatment (χ²=0.134, P=0.714). Conclusions The Wumeiwan combined with triple therapy for treatment of CAG has a good efficacy and low adverse reactions, has serious anti-inflammatory effects, can improve the body immunity, and it was worthy clinical application.

OBJECTIVE: To investigate the expression and clinical pathological significance of PDCD4 in laryngocarcinoma tissue and its potential significance to clinic. METHOD: Western-blotting and immunohistochemistry ana lyse to measure the protein expression of PDCD4 in 54 cases of laryngocarcinoma tissues (studying group) and their paraneoplastic normal tissues (control group). The correlations of PDCD4 with clinical pathological parameters were analyzed. RESULT: PDCD4 protein was positively expressed in paraneoplastic normal tissue while which was lost or decreased in laryngocarcinoma tissue by Western blot and immunohistochemistry. Immunohistochemistry assay showed the location of PDCD4 protein in cells was different between the studying group and the control group. The expression level of PDCD4 was related to the pathological grades of the laryngocarcinoma. It's higher in the well-differentiated tumor group than that in the poorly differentiated ones. But the expressions of PDCD4 were no differences among other clinical parameters including sex, age, clinical classification, clinical stage and the cervical lymphonodus who had been metastases or not. CONCLUSION PDCD4 gene is anti-oncogene. It may play an important role in the pathogenesis and development of laryngeal carcinoma and it may be a new target of therapy for laryngo carcinoma.
Adult ; Aged ; Aged, 80 and over ; Apoptosis Regulatory Proteins ; metabolism ; Female ; Genes, Tumor Suppressor ; Humans ; Immunohistochemistry ; Laryngeal Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; RNA-Binding Proteins ; metabolism

Objective To study the developmental process of the region of basal nuclei of postmortem fetuses by 3.0 T and 7.0 T MRI.Methods One hundred and thirty-one postmortem fetuses of 14 to 40 weeks of gestational age(GA)were scanned by 3.0 T MR,of which 11 fetuses of 14-27 weeks of GA were chosen and scanned by 7.0 T MR. The time when the structures in the region of basal nuclei could be detected and the changes of MR signal intensity were analyzed for MRI of different Tesla.Results On 3.0 T MRI.the dorsal thalamus could be delineated as early as 14 weeks of GA. The germinal matrix, caudate nucleus,and putamen could be visualized as early as 15 weeks of GA. The globus pallidus could be described as early as 18 weeks of GA.and the internal capsule and external capsule could be shown as early as 20 weeks of GA. The signal of the caudate nucleus during 15-30 weeks of GA was relatively hypointense on T1WI and hyperintense on T2WI.but during 31-40 weeks of GA, it was relatively hyperintense on T1WI and hypointense on T2WI. The putamen had a relatively high signal intensity on T1WI and low signal intemity on T1WI during 15-17 weeks of GA, and it appeared patchy during 18-25 weeks of GA,then it had a relatively low signal intensity on T1WI and high signal intensity on T2WI during 26-30 weeks of GA, and during 31-40 weeks of GA,its signal intensity was relatively high on T1WI and low on T2WI.The globus pallidus had a relatively high signal intensity on T1WI and low signal intensity on T2WI during 20-40 weeks of GA Compared to the 3.0 T MRI,the T2 images of 7.0 T MRl were more clear,and most structures in the region of basal nuclei could be clearly displayed as early as 16 weeks of GA.such as the germinal matrix,caudate nucleus,dorsal thalamus,putamen,globus pallidus,internal capsule,and external capsule.The claustrum could be delineated as early as 18 weeks of GA on 7.0 T MRI. Conclusions 3.0 T MRI could show the developmental process of the region of basal nuclei well,but the T2 images of 7.0 TMRl were comparatively better.

Objective To clone and identify the differentially expressed genes of rat intestinal epithelial cell line (IEC 6) before and after exposure to high dose radiation so as to provide proof for the investigation of the molecular mechanisms in the repair of radiation damage of intestinal epithelial cells. Methods A subtractive cDNA library for differentially expressed genes was constructed by suppression subtractive hybridization (SSH) and T/A cloning technique after IEC 6 cells were exposed to radiation at the dose of 35 Gy ? ray. The expressed sequence tag (EST) library was screened by reverse Northern hybridization. Positive clones were sequenced and the similarity was searched against the DNA database in GenBank. Limited clones were identified by Northern hybridization. Results More than 2 000 white clones were harvested after the library amplification. Ninety six of them were randomly picked out for PCR amplification, and 15 positive clones which corresponded to 12 individual genes were identified by reverse Northern hybridization. These genes were involved in cell skeleton, cell stress, cell cycle control, and signal transduction, etc. In addition, a novel cDNA sequence was also obtained. Conclusion A subtractive cDNA library for differentially expressed genes in IEC 6 cells exposed to the radiation at the dose of 35 Gy ? ray has been successfully constructed with SSH and T/A clone techniques. Several positive ESTs which correspond to genes involving in cell skeleton, cell stress, cell cycle control, and signal transduction are identified. These genes may play important roles in the process of the damage and repair of the intestinal epithelial cells exposed to radiation.

BACKGROUND: The hepatitis B virus (HBV) vaccine has been efficiently used for decades. However, hepatocellular carcinoma caused by HBV is still prevalent globally. We previously reported that interferon (IFN)-induced tripartite motif-containing 25 (TRIM25) inhibited HBV replication by increasing the IFN expression, and this study aimed to further clarify the anti-HBV mechanism of TRIM25. METHODS: The TRIM25-mediated degradation of hepatitis B virus X (HBx) protein was determined by detecting the expression of HBx in TRIM25-overexpressed or knocked-out HepG2 or HepG2-NTCP cells via Western blotting. Co-immunoprecipitation was performed to confirm the interaction between TRIM25 and HBx, and colocalization of TRIM25 and HBx was identified via immunofluorescence; HBV e-antigen and HBV surface antigen were qualified by using an enzyme-linked immunosorbent assay (ELISA) kit from Kehua Biotech. TRIM25 mRNA, pregenomic RNA (pgRNA), and HBV DNA were detected by quantitative real-time polymerase chain reaction. The retinoic acid-inducible gene I (RIG-I) and pgRNA interaction was verified by RNA-binding protein immunoprecipitation assay. RESULTS: We found that TRIM25 promoted HBx degradation, and confirmed that TRIM25 could enhance the K90-site ubiquitination of HBx as well as promote HBx degradation by the proteasome pathway. Interestingly, apart from the Really Interesting New Gene (RING) domain, the SPRY domain of TRIM25 was also indispensable for HBx degradation. In addition, we found that the expression of TRIM25 increased the recognition of HBV pgRNA by interacting with RIG-I, which further increased the IFN production, and SPRY, but not the RING domain is critical in this process. CONCLUSIONS The study found that TRIM25 interacted with HBx and promoted HBx-K90-site ubiquitination, which led to HBx degradation. On the other hand, TRIM25 may function as an adaptor, which enhanced the recognition of pgRNA by RIG-I, thereby further promoting IFN production. Our study can contribute to a better understanding of host-virus interaction.
Humans ; Hepatitis B virus ; DEAD Box Protein 58/metabolism* ; RNA ; Liver Neoplasms ; Virus Replication ; Tripartite Motif Proteins/genetics* ; Transcription Factors ; Ubiquitin-Protein Ligases/genetics*