With the economic growth and better standards of living,the prevalence of non-alcoholic fatty liver disease (NAFLD) is expected to increase dramatically worldwide.NAFLD is a common chronic inflammation disease.NF-κB is a transcription factor that plays crucial roles in inflammation,immunity,cell proliferation and apoptosis.It can facilitate the occurrence and development of NAFLD,and the underlying mechanisms are related to insulin resistance,oxidative stress,alteration of intestinal flora,activation ofrenin angiotensin system,etc.

Objective To evaluate the in vitro inhibiting activities of interferon-alpha (INF-?) against hepatitis B virus (HBV). Methods HepG2 2 2 15 cells were treated with INF-?. At 3rd, 6th and 9th days after treatment, the supernatant was collected for HBsAg quantitative assay, and total RNA of the cells was isolated for RT-PCR and realtime-PCR assay of HBV mRNA. Results There were no significant differences in HBsAg titer and virus mRNA level at 3rd and 6th days between the experimental and control groups. At 9th day, HBsAg titer and virus mRNA level in INF-? group were significantly lower than those in control group. And 500IU/ml INF-? could get maximal inhibiting effect against HBV. Conclusion INF-? can inhibit transcription and expression of HBV gene in HepG2 2 2 15 cells, which indicated that INF-? can inhibit HBV replication directly.

Objective To determine the changes of serum glycosylphosphatidylinositol specific phospholipase D(GPI-PLD) activity in patients of several liver disease.Method Glycosylphosphatidylinositol (GPI) anchored placental alkaline phosphatase(PLAP) prepared by ourselves was used as a substrate.After partitioning by triton-X-114,the serum GPI-PLD activity was determined quantitatively and the data was treated by microware of SPSS 10.0.Results On the basis of the percentage of GPI-anchored PLAP conversion,the sera GPI-PLD activities of total 172 patients,included 26 severe acute viral hepatitis as group A,29 liver cirrhosis as group B,32 chronic viral hepatitis as group C,55 mild acute viral hepatitis as group D,30 primary hepatocellular carcinoma as group E,were measured.As compared with 182 healthy presons as control group,the sera GPI-PLD activities of group A and B were significantly reduced;By contraries,the activities of group D and E were significantly raised.The sera GPI-PLD activities of group C compared with healthy control group were not significantly altered.However,when paired Q-test,the changes of serum GPI-PLD activity between all paired groups among this five groups were remarkable.Conclusions The determination of sera GPI-PLD activities in patients can act as a biochemistry index for diagnosis of acute viral hepatitis,liver cirrhosis and primary hepatocellular carcinoma,as well as an auxiliary index for judgment of the curative effect and prognosis of liver diseases.

Objective To evaluate the in vitro inhibitive activities of mycophenolic acid against hepatitis B virus in vitro. Methods The different concentrations of the test drugs in DMEM were added to the well with the monolayer growth of HepG2.2.15 cells. The culture medium was refreshed with DMEM containing the appropriate drugs every 3 days. At 3rd, 6th and 9th days, the supernatant was collected for HBsAg quantitative assay, total RNA was isolated from cells for RT-PCR and realtime-PCR assays. Results It was found no significant anti-HBV effect of mycophenolic acid on concentration of 2～250 ?g/ml, and mini inhibitory efficacy of HBV replication on concentration of 1250 ?g/ml. There was significant anti-HBV effect of lamivudine on concentration of 5 ?g/ml. When combined with lamivudine mycophenolic acid can significantly potentiate anti-HBV activities of lamivudine, and it can get maximal inhibition effect on concentration of 10 ?g/ml. Conclusions Mycophenolic acid could inhibit HBV replication and potentiate anti-HBV activities of lamivudine.

Objective To investigate gene variation and the relationship between gene variation and pathogenicity of transfusion transmitted virus(TTV).Methods The TTV DNA in the serum sample from a blood donor(BD) and a chronic non-A-G severe hepatitis(CSH) patient with TTV infection was amplified by using PCR.The purified PCR product was cloned and 10 clones from each case were sequenced.The sequences were compared among different clones and analyzed by Phylogentic tree.Results There were two different TTV strains in the BD and seven different TTV strains in the chronic non-A-G severe hepatitis patient.The TTV clones in the BD were of G1a subtype and those of the CSH were of G1a and G1b subtype.Conclusion Gene variant of TTV was much more complicated in the CSH patients than that in the BD ones.

arkably increases with the development of cirrhosis,which may play an important role in the development of PHG.AG may remarkably ameliorate the degree of PHG，mainly by inhibiting the expression of iNOS in gastric musosa.

Objective To investigate the effect of exogenous vascular endothelial growth factor (VEGF) on bone activity of rabbit heterotopic allograft decalcified bone.Methods 140 adult healthy China white rabbits were selected,no limitation with sex,20 rabbits as the donor preparation of allogenic decalcified bone,according to the random number table,the rest was divided into the experimental group (allograft decalcified bone ± VEGF) and the control group (Allograft decalcified bone),each group contained 60 rabbits.For the experimental group,the prepared 1.5 cm long homologous decalcified tibia was placed in rabbit right thigh of rectus femoris and vastus medialis muscle gap near by saphenous artery,and fixed on the femur with two 0.8 mm Kirschner wire.In the vicinity of the skin,implanted an osmotic pump which contain the VEGF solution 200 μl with concentration was 0.5 μg/ml.In the control group,implanted the isometric allograft decalcified bone in rabbit right thigh corresponding parts with the same method.Each group respectively at 0,2,4,6,8,10 weeks to death 10 white rabbits,By specimen observation,HE dyeing observation and detection of type Ⅰ glue protein fluorescence intensity,Analysis the bone activation degree of two groups of bone allograft decalcified.Results Experimental allograft decalcified bone gradually wrapped by connective tissue membrane,its surface appear different size of the pits and gradually increased and become deep,while the control group pits relatively little and shallow.In the experimental group and control group,the fluorescence intensity of type Ⅰ collagen reached its peak respectively at 8 weeks (47.57 ±3.50) and 10 weeks (45.07±6.02),with no statistically significant (P ＞ 0.05).Conclusion Rabbit allograft decalcified bone implanted in the muscle clearance with abundant blood supply can be transformed into activated bone after 10 weeks,and after applying exogenous VEGF,allograft decalcified bone can be transformed into activated bone after 8 weeks,the bone activation process obviously speed up.The reaults confirmed the exogenous VEGF can obviously promote the ectopic rabbit bone allograft decalcified bone activation process.

Objective To investigate clinical features and antifungal therapeutic effect of chronic severe hepatitis (CSH) patients with invasive fungal infection (IFI), and to improve the diagnosis and treatment.Methods Clinical manifestation, blood routine, imageology and mycetology characteristic, antifungal treatment perscription and therapeutic effect of 79 CSH patients with IFI were retrospectively analyzed. Antifungal therapeutic effect was compared between fluconazole and voriconazole. Results Thirteen (16.5%) patients received glucocorticoid or other immunodepressants for a relatively long time, 40 (50.6%) patients had invasive operation, and 61 (77.2 %) patients were administered 1-6 kinds of broad-spectrum antibiotics. Seventy-three patients had fever. Leucocytes and neutrophilic granulocyte increased in 96.2% of the patients. Lung (31.6%), intestinal tract (26.2%) and oral cavity (14%) infections were common. Fungus was found in 70.9% of the patients. Candida albicans (40.9%) and aspergillus (21.1%) were often seen. Halo signs and crescent signs on lung CT were relatively specific in 40% of the patients with fungal pneumonia. Voriconazole was more effective than fluconazole(71.4% vs. 39.0%, P<0.05). Twelve patients with lung aspergillus infection were administered voriconazole, 8 (66.7%) patients of whom was effective, and the other 4 (33.3%) patients died. Conclusion There are high risk factors in major CSH patients with IFI. The most common clinical manifestations of CSH patients with IFI are fever, leukocytosis, lung and intestinal tract infection. Candida albicans and aspergillus infection are common. Voriconazole is more effective than fluconazole, and can increase the survival rate of CSH patients with IFI.

Objective To investigate the effect of erythropoietin(EPO)on the expression of aquaporin - 2 (AQP2)in the kidney of young SD rats after release of bilateral ureter obstruction(BUO - R). Methods Thirty - two young SD rats were equally divided into 4 groups randomly(BUO group,BUO - R group,BUO - R ﹢ EPO group and Sham group,8 rats in each group). The BUO model was built through bilateral ureteral ligation. EPO(500 U/ kg)was given to BUO - R ﹢ EPO rats at 2 h after release of BUO,and then repeated 6 h,12 h,24 h and 36 h thereafter and the same volume of 9 g/ L saline was simultaneously given to BUO - R rats. The Sham group was prepared in parallel by laparotomy and free dissection of bilateral ureters but not ligated. Both side kidneys were harvested 48 h(72 h for Sham group)after release of BUO to examine the effect of EPO on the expression of AQP2 in inner medulla by immunohisto-chemistry,Real - time PCR and Western blot. The urine samples were collected by using metabolic cage before death. Results The osmotic pressure of BUO - R ﹢ EPO group was higher than that of BUO - R group,but lower than that of Sham group(P ﹤ 0. 05). Immunohistochemistry showed that the collecting duct wall thinned and lumen enlarged. After the pictures were analysized by using Image - Pro Plus software,it showed that the expression of AQP2 in collecting duct in BUO group was significantly down - regulated compared with that in Sham group,whereas,it was slightly weaker in BUO - R group and BUO - R ﹢ EPO group than Sham group(P ﹤ 0. 05). These results were further confirmed by a-dopting Western blot,and the relative quantity of AQP2 in BUO group was also the lowest of the four groups(P ﹤0. 05). Real - time PCR showed that the level of AQP2 mRNA in Sham group was(24. 30 ± 1. 03)folds of BUO group,(10. 60 ± 1. 05)folds of BUO - R group and(5. 70 ± 1. 01)folds of BUO - R ﹢ EPO group,respectively. Conclusion EPO could promote not only the recovery of AQP2 mRNA and protein expression but also the recovery of AQP2 function in young BUO - R rats.

OBJECTIVE: To screen differentially expressed genes and gene pathways in L02 cell line stably expressing hepatitis C virus (HCV) Ib type nonstructural protein 4B (NS4B) mediated by lentiviral system, and to provide a basis for further research of molecular biological mechanism of NS4B gene in chronic hepatitis C and hepatocarcinogenesis. METHODS: NS4B stably overexpressed L02 cell line and negative control stable L02 cell line, designated as L02-NS4B and L02-mkate2 respectively, were resurrected and amplified in vitro. The differentially expressed genes between L02-NS4B and L02-mkate2 were determined by gene expression microarray from Human Gene 1.0ST. The significant pathways of the differential genes were selected by the Fisher's exact test and χ2 test according to kyoto encyclopedia of genes and genomes (KEGG) database. The differential expression levels of 5 selected genes including protein kinase C delta binding protein (PRKCDBP), tumor protein p53 (TP53), v-akt murine thymoma viral oncogene homolog 1 (AKT1), baculoviral IAP repeat containing 3 (BIRC3) and B-cell lymphoma 2-like1 (BCL2L1) from cDNA microarray data were further verified by real-time quantitative polymerase chain reaction (real-time QPCR). RESULTS: Between L02-NS4B and L02-mkate2, the genes with flurescence intensity ratio >1.2 or <0.8 were considered as differentially expressed genes. A total of 2 682 differentially expressed genes in the known 28 869 human genes were detected in L02-NS4B, 1 446 genes were upregulated and 1 236 genes were downregulated. A total of 41 involved pathways of up-regulated differential genes were identified by KEGG database, mainly including apoptosis, extracellular matrix receptor interaction, cell cycle, pathways in cancer and Toll-like receptor signaling pathway; and 20 involved pathways of down-regulated differential genes were identified, mainly including pathways in cancer, Wnt signaling pathway and cell cycle pathway. Of the 5 upregulated genes selected from cDNA microarray data, 3 genes showed the same differential expression pattern by real-time QPCR as that shown in cDNA microarray data, namely AKT1, BIRC3 and BCL2L1. The confirmation rate of real-time QPCR was 60%. CONCLUSION HCVNS4B can up-regulate or down-regulate the expression of many genes in L02 cells, thus affecting multiple signaling pathways relevant to cell apoptosis, cell cycle and carcinogenesis.
Cell Cycle ; Cell Line ; Gene Expression Profiling ; Hepacivirus ; genetics ; Humans ; Oligonucleotide Array Sequence Analysis ; Real-Time Polymerase Chain Reaction ; Signal Transduction ; Transcriptional Activation ; Viral Nonstructural Proteins ; genetics