The natural history of chronic HBV infection is diverse and variable, ranging from inactive carriers to progressive chronic hepatitis B ( CHB), cirrhosis and hepatocelluar carcinoma.It is estimated that 93 million people are chronically infected with HBV and 20 million cases suffering from chronic hepatitis B in China.Hepatocelluar carcinoma has been the second leading cause of death for male in China.Liver cirrhosis and HCC which have high mortality and morbidity have become the heavy burden for the limited medication resource of China.Here the current clinical applications and consensus progression based on antigen and nuclear acid detection were acknowledged.The reasonable application as well as appropriate clinical interpretation are emphasized indicating that laboratory medicine practitioners should be more actively involved in clinical diagnosis and treatment.More efforts and contributions should be made by the laboratory medicine practitioner for optimizing clinical management of HBV-related diseases in future.

China is among the middle-high endemic regions of HBV infection.The pathological outcomes of chronic HBV infection have been shown to be greatly influenced by several important factors,including HBV genotype,sub-genotype and gene viability mutation.HBV genome mutation,on the one hand,could alter its replication and secretion and thus change viral pathogenicity.In addition,host immune microenvironment and host-virus interaction,disease progression and the effect of antiviral therapy could be adapted at the same time.The detection of HBV genotypes,genetic subtypes and the key hotspot mutation is helpful to clinical risk assessment and prognosis prediction of HBV-related end-stage liver diseases (cirrhosis and hepatocellular carcinoma),it is also helpful to auxiliary predict the liver diseases recurrence and metastasis after treatment.Thus persistent care should be taken on the HBV mutation and its clinical translation so as to provide solid evidences for the personalized,standardized and fine management of HBV-related liver diseases.

Objective To investigate the oxidative mechanism of homocysteine ( Hey) -induced apoptosis in endothelial progenitor cells( EPCs). Methods Total mononuclear cells were isolated from mouse bone marrow by Ficoll density gradient centrifugation and were cultured in vitro for 7 d. Adherent cells were harvested and identified by fluorescence microscopy. EPCs were cultured with Hey (0, 50, 100 and 500 μmol/L) for 12, 24 and 48 h, or pretreated with NAC (1 mmol/L), DPI( 10 μmol/L) or SB203580 (10 μmol/L) for 30 min, then cultured with 500 μmol/L Hey for 24 h. Apoptosis was detected by Annexin-V/PI flow cytometry, levels of reactive oxygen species (ROS) in cells were measured using H2DCF-DA as a fluorescence probe, NADPH oxidases were evaluated with lucigenin-enhanced chemilumine9cence, and NO in the supernatant was determined by nitrate reductase assay. Results Hey induced EPCs apoptosis, ROS accumulation, NADPH oxidase activation and decrease of NO in a time-dose dependent manner( P <0.05 or P < 0.01). Pretreatment with NAC, DPI and SB203580 could inhibit these effects (P < 0.05 or P < 0.01). Conclusion Hey could activate NADPH oxidase, induce ROS increase and NO decrease, and activate p38MAPK to enhance EPCs apoptosis.

Objective To observe the expression and distribution of IL 10 in local vessel after balloon injury in rat and study its significance in the response of artery injury Methods RT PCR and Western Blot as well as immunohistochemical method were used to assay the changes of IL 10′s mRNA as well as expression and distribution of protein Results There is no expression of IL 10 in vascular wall of normal rat After Balloon injury, the level of IL 10 mRNA and product of IL 10 was up regulated, and immunohistochemiscal staining showed immunoreactive IL 10 mainly in smooth muscle cells Conclusion IL 10 is expressed in balloon injured aorta and may contribute to the modulation of the local inflammatory response

To evaluate the feasibility of using polyethyleneimine (PEI) coated magnetic iron.oxide nanoparticles (polyMAG-1000) as gene vectors. The surface characteristics of the nanoparticles were observed with scanning electron microscopy. The ability of the nanoparticles to combine with and protect DNA was investigated at different PH values after polyMAG-1000 and DNA were combined in different ratios. The nanoparticles were tested as gene vectors with in vitro transfection models. Under the scanning electron microscope the nanoparticles were about 100 nm in diameter. The nanoparticles could bind and condense DNA under acid, neutral and alkaline conditions, and they could transfer genes into cells and express green fluorescent proteins (GFP). The transfection efficiency was highest (51%) when the ratio of nanoparticles to DNA was 1:1 (v:w). In that ratio, the difference in transfection efficiency was marked depending on whether a magnetic field was present or not: about 10% when it was absent but 51% when it was present. The magnetic iron oxide nanoparticles coated with PEI may potentially be used as gene vectors.
Breast Neoplasms/metabolism ; Breast Neoplasms/*pathology ; Cell Line, Tumor ; Ferric Compounds/*chemistry ; Ferric Compounds/metabolism ; Gene Targeting ; Genetic Vectors ; Green Fluorescent Proteins ; Magnetics ; Nanotechnology ; Particle Size ; Polyethyleneimine/chemistry ; Transfection/methods

The aim of this study was to evaluate the feasibility and efficacy of using TRAIL gene to treat breast cancer mediated with a novel carrier - magnetic iron oxide nanoparticles (poly-MAG-1000) coated with PEI. The magnetic iron oxide nanoparticles were used as gene carrier to transfect TRAIL gene into MCF-7 cells. The polyMAG-1000 without TRAIL gene was transfected into the tumor cells as negative control. TRAIL gene transfection with liposome as carrier served as positive control. The apoptosis of cells was detected with TUNEL method. The apoptosis ratio of tumor cells was measured with flow cytometry (FCM). It was found that the apoptosis occurred in the tumor cells after transfection of TRAIL gene mediated by both polyMAG-1000 and liposome. The apoptosis ratio in the group with polyMAG-1000 as gene carrier was (25.11+/-2.85) %, whereas it was (5.06+/- 1.05) % in the control group with polyMAG-1000 (P<0.01). The apoptosis ratio was as low as (18.31+/-2.44) % in the group with liposome as gene carrier (P<0.05, as compared with the group with polyMAG-1000 as gene carrier). It is suggested that TRAIL gene may induce apoptosis in MCF-7 breast cancer cells. The magnetic iron oxide nanoparticles coated with PEI may be a potential gene carrier with high transfection efficacy for cancer gene therapy..

Objective To investigate the therapeutic effect of frozen-thawed murine islets which were transplanted into diabetic rats after cultured with hyperbaric oxygenated rotary cell culture system (HORCCS). Methods The purified rat islets were divided into two groups: A. In vitro experiment groups (IvEG) : The rat islets in each subgroup were cultured in HORCCS or common medium for 30 days, then evaluated for the intracellular DNA and insulin contents of islets, and the viability and insulin secreting level of islets. B. Islet transplantation experimental groups (TxEG) : The frozen-thawed islets were cultured in HORCCS or common medium for 7 days, and then transplanted into the recipients. We observed the blood glucose level (BGL) and insulin secreting level in the recipients as well as the uhrastructure change of islets in TxEG. Results The viability and insulin secreting level of islets cultured with HORCCS at 14th day were much higher than those cultured with common medium (P <0.05). The blood glucose level in recipients transplanted with islets cultured with HORCCS recovered to normal value at the 2nd week and lasted for 8 weeks. All these recipients maintained the normal glucose tolerance curve. Electronic microscopy found microchannel outlets on the surface of the frozen-thawed islets cultured with HORCCS. Conclusions Frozen-thawed islets cultured with HORCCS could establish nutrient transmission microchannels, which were not only capable of oxygen and nutrients transmission, but also improving cryopreservation solution to diffuse inside the islet cells evenly and uniformly. So this method not only lessens islet damage from cryopreservation, but also improves the effect of transplantation.

Aim To study the effect of TSA on vascular smooth muscle cells(VSMC)proliferation and apoptosis in vitro.Methods VSMC proliferation was analyzed by MTT assay and BrdU incorporation assay.Cell cycle phase distributions were determined by flow cytometer.The expressions of cyclin D1 and cyclin A were assessed by western blot.Cell apoptosis was quantified by detecting cytoplasmic histone-associated DNA-fragments and the level of cleaved caspase-3.Results TSA at a low concentration was adequate to inhibit serum-induced VSMC proliferation without significant cytotoxity.High concentration of TSA activated caspase-3 and induced VSMC apoptosis.TSA treatment reduced expressions of cyclin D1 and cyclin A,and blocked VSMC entry into S phase.Conclusions TSA inhibits serum-induced VSMC proliferation and G1→S phase progression of cell cycle.Histone deacetylase(HADC)inhibitors may constitute a novel therapy for vascular proliferative diseases.

Objective:To investigate the effects of OX-LDL on the formation and progression of atherosclerosis. Methods: (1) The thoracic aorta smooth muscle cells(VSMC) of male Sprague-Dawley rat were primarily cultured in vitro. Different concentrations of OX-LDL(50,100,150,200 ?g/ml) were co-incubated with the cells respectively. MTT method was used to detect the cell proliferation after 24, 48 and 72 h. (2) pCOLH22. 4 and pCOLH2l. 6, recombinants of human a2( I ) procol-lagen gene 5' flank sequence ( - 2. 4 kb and -1. 6 kb in size) and CAT reporter gene, were used to transiently transfect smooth muscle cells with FuGENE Transfectant Reagent. The effect of OX-LDL (150 ?g/ml) on the plasmid were determined by CAT-EL1SA. Results: (1)OX-LDL accelerated the proliferation of VSMC in dose-dependent manner. (2)OX-LDL enhanced the promoter activities of human a2( I ) procollagen gene significantly. Conclusion: OX-LDL can accelerate the proliferation of VSMC and the formation of type I collagen,which facilitates the formation and progression of atherosclerosis.

AIM:To investigate the effect of trichostatin A(TSA)on p27kip1 gene expression in vascular smooth muscle cells.METHODS:Reverse transcription-polymerase chain reaction(RT-PCR)was used to measure the level of p27kip1 mRNA.The protein levels of p27kip1 and S-phase kinase-associated protein-2(skp2)were determined by Western blotting.20S proteasome activity was quantified by using a fluorogenic proteasome-specific substrate.RESULTS:TSA did not affect mRNA level of p27kip1 in VSMCs,but attenuated serum-induced downregulation of p27kip1 through stabilizing p27kip1 turnover.In addition,TSA decreased the expression of skp2,an F-box protein that targets p27kip1 for degradation,but had no effect on proteasome activity.CONCLUSION:TSA regulates p27kip1 expression at the post-translational level in VSMCs.