Animals ; Hepacivirus ; genetics ; Hepatitis B ; therapy ; Hepatitis B virus ; genetics ; Hepatitis C ; therapy ; Humans ; Mice ; RNA Interference ; RNA, Small Interfering ; therapeutic use ; Virus Replication

Antiviral Agents ; therapeutic use ; Hepatitis C, Chronic ; drug therapy ; Humans ; Interferons ; therapeutic use

Objective To investigate the correlation between ~(18)F-fluorodeoxyglucose (FDG) uptake and hypoxia inducible factor1α (HIF-1α) level,microvessel density (MVD) in human gliomas.Methods ~(18)F-FDG PET scan was performed preoperatively in 41 patients with gliomas (including 23 highgrade and 18 low-grade tumors).The ratios of maximum standardized uptake value(SUV_(max))between tumor (T)and contralateral white matter (WM) were calculated (T/WM).Immunohistochemical stain methods were used to evaluate the level of HIF-1α and measure the MVD in tumors.Correlation analysis between SUV_(max) of T/WM and HIF-1α level,MVD wag performed.The t-test,one-way ANOVA test,Spearman rank correlation and Wilcoxon signed-rank test were calculated using SPSS 11.5 software.Results (1)The SUV_(max) of T/WM,HIF-1α level and MVD in high-grade and low-grade tumors groups were 3.39±1.43,95.7% and 44.13±16.1 vs 1.46±0.55.55.6% and 18.83±7.07,respectively.The difierences of SUV_(max) of T/WM,HIF-1α level and MVD between two groups were statistically significant (t=-5.921,z=-3.938,t=-6.745,all P＜0.05).(2)Among 41 gliomas,the strong positive expression of HIF-1α was observed in 8,mederate in 9,weak in 15 and negative expression was found in 9,SUV_(max) of T/WM and MVD increased with increasing HIF-1α level.The differences of SUV_(max) of T/WM and MVD among 4 different groups were statistically significant (F=7.41,P＜0.05).(3) The MVD of all gliomas was ranged from 9.76 to 94.52,which correlated with SUV_(max) of T/WM(r=0.759,P＜0.05).Conclusions The SUV_(max) of T/WM correlates with HIF-1α level and MVD in gliomas.Therefore,~(18)F-FDG PET provides preoperatively a noninvasive assessment of hypoxia or angiogenesis in human glionma.

OBJECTIVESBased on the immunologic character of Pichia pastoris yeast, a new therapeutic vaccine, whole recombinant yeast, was used to explore a new way to activate cell-mediated anti-viral immunity.

METHODSThe recombinant plasmids, pPIC9K/S and PIC9K/hsp(1-370)-S, were constructed by inserting the gene encoding HBsAg, HSP70 (1-370) -HBsAg into vector pPIC9K and then the recombinants were transfected into Pichia pastoris yeast,GS115, respectively. Then that recombinant yeast immunized BALB/C mice were detected for humoral and cellular immunity to HBsAg.

RESULTSRecombinant yeast successfully activated the humoral immunity to HBsAg in mice, but failed to activate the cellular immunity.

CONCLUSIONThe whole recombinant yeast can be used as vaccine, but need further study for optimal way of immunization.

Animals ; Female ; HSP70 Heat-Shock Proteins ; genetics ; immunology ; Hepatitis B Antibodies ; blood ; Hepatitis B Surface Antigens ; genetics ; immunology ; Hepatitis B Vaccines ; immunology ; Mice ; Mice, Inbred BALB C ; Pichia ; genetics ; Plasmids ; T-Lymphocytes, Cytotoxic ; immunology ; Vaccines, Synthetic ; immunology

OBJECTIVETo identify the siRNA interference ability for the replication of HBV.

METHODSBased on the sequence of HBV in HepG2 2.2.15 cells in GenBank, one sequence targeting the C antigen of HBV was cloned into the RNA polymerase III based expression vector pSuper. This recombinant was electroporated into HepG2 2.2.15 cells and the expression of HBsAg and HBeAg was assayed using ELISA.

RESULTSThe construction of the recombinant expression vector pSuper-C and its control vector pSuper was successfully confirmed by the results of enzyme digestion, electrophoresis and sequencing. However, there was no difference between the expression of HBsAg and HBeAg in the supernatant of HepG2 2.2.15 cell culture in the experimental and control groups.

CONCLUSIONSThe constructed pSuper-C did not show an interfering effect on the replication of HBV in HepG2 2.2.15 cells. In order to display this effect, further study is needed

Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; virology ; RNA Interference ; RNA, Small Interfering ; genetics ; Tumor Cells, Cultured ; Virus Replication ; genetics

During the past 70 years since the founding of New China, Chinese public health especially the prevention and control of infectious diseases have made remarkable achievements, which benefited from the vaccination. This study is to summarize the progress of immunization and vaccines, the achievements and contributions of vaccines including polio vaccine, hepatitis B vaccine, diphtheria, tetanus and acellular pertussis combined vaccine, measles vaccine and hepatitis A vaccine to the prevention and control of infectious diseases in China in the past 70 years and to review the research and development of innovative vaccines in China in recent years, which may provide clues for the development of the expanded programe on immunization in China in the future.

OBJECTIVERegarding the strong antigen-presenting abilities of dendritic cells (DC), this study was carried out based on the induction and proliferation of DC derived from human umbilical cord blood; the anti-HBV effect of cytotoxicity T lymphocytes (CTL) activated by those DC pulsed with HBsAg was also carried out to explore a new way to activate the HBsAg-specific CTL.

METHODSCord blood was collected from the cord veins of normal placentae after Cesarean sections, from which cord blood mononuclear cells (CBMC) were separated through density gradient centrifugation. The CBMC were cultured in RPMI 1640 with a cytokine cocktail. Pulsed with HBsAg, the DC were prepared to activate the HBsAg-specific CTL among the CBMC. The cytotoxic effect of CBMCs activated by the DC primed with HBsAg was assayed through the killing of those HepG2-S target cells.

RESULTSTypical DC could be induced from CMBC cultured with a cytokine cocktail. DC pulsed by HBsAg activated HBsAg-specific CTL, which killed the target HepG2-S cells to some extent.

CONCLUSIONDC can be induced from CMBC with the cytokine cocktail and they show a strong antigen-presenting ability. DC produced in this way and pulsed by HBsAg can activate HBsAg-specific CTL in vitro. This might mean that it could be a new way to break the tolerance to HBV in chronic HBV-infected patients.

Cell Culture Techniques ; Cells, Cultured ; Dendritic Cells ; cytology ; immunology ; Fetal Blood ; cytology ; Hep G2 Cells ; Hepatitis B Surface Antigens ; immunology ; Hepatitis B virus ; immunology ; Humans ; T-Lymphocytes, Cytotoxic ; immunology

Dendritic cell (DC) plays a key role in antitumor immune response. However, there is a deficiency of DC function in the majority of leukemia patients. It is a novel idea that expanding DC in vitro and enhancing their antitumor immune function and DC-based tumor vaccines may be used as an efficient immune therapy for leukemia. In the project, the condition to induce DC from myeloid leukemia cell lines and its anti-leukemia response were investigated. HL-60, K562 and THP-1 cells were cultured with various combinations of cytokines for inducing DC. The morphologic features were analyzed with optical and electron microscopy. The phenotype of DC was detected by FCM with CD1a, CD40, CD80, CD86, HLA-A, B, C and HLA-DR monoclonal antibodies. The ability of DC stimulating lymphocyte proliferation was observed by allo-mixed lymphocyte reaction using (3)H-TdR incorporation. Cytotoxicity assay was measured by (51)Cr-release method. The level of IL-12 and IFN-gamma in supernatant of DC culture was measured by ELISA. It was proved that the DCs derived from K562, HL-60 and THP-1 cells showed a typical morphology of dendritic cell. The induced cells expressed the surface differentiation antigens of DC. A high expression of phenotypes was found in HL-60-DC and THP-1-DC stimulated by GM-CSF + IL-4 + TNF-gamma and K562-DC with GM-CSF + IL-4 + IL-12. The DCs from the 3 leukemia cell lines stimulated allo-MLR and CTL reaction strongly. Different contents of IL-12 were detected in the supernatants of DC culture and IFN-gamma in the coculture of DC and blood mononuclear cells. It is concluded that the myeloid leukemia cells are able to be induced DCs by cytokines in vitro. The different leukemia cells need different cytokines and cultural conditions. DCs derived from leukemia cells express phenotype of antigen-presenting cells. They have the ability of stimulating T lymphocyte proliferation and inducing CTL reaction to clear leukemia cells, and the DCs secrete IL-12 and increase secretion of IFN-gamma by T cells.
Antigens, CD ; immunology ; Antigens, CD1 ; immunology ; B7-2 Antigen ; CD40 Antigens ; immunology ; Coculture Techniques ; Cytokines ; pharmacology ; Cytotoxicity, Immunologic ; drug effects ; Dendritic Cells ; drug effects ; immunology ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; HL-60 Cells ; HLA Antigens ; immunology ; Humans ; Immunophenotyping ; Interferon-gamma ; pharmacology ; Interleukin-12 ; pharmacology ; Interleukin-4 ; pharmacology ; K562 Cells ; Membrane Glycoproteins ; immunology ; Microscopy, Electron ; Time Factors ; Tumor Cells, Cultured ; drug effects ; immunology ; ultrastructure ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo investigate transient expression of fusion protein with a chimeric HBsAg-HSP70 construct in HepG2 cells.

METHODSEnkaryotic expression plasmids inserted HBsAg gene or chimeric HBsAg-HSP70 gene were prepared and transfected into HepG2 cells by means of cationic liposome. mRNA were detected by RT-PCR and proteins expressed in the cells were detected by immunocytochemistry 48 hours later. HBsAg in cultured supernatants and cell lysates were assayed by ELISA.

RESULTSFusion protein (HBsAg-HSP70) transient expression in HepG2 cells were confirmed by RT-PCR, immunocytochemistry or ELISA, but fusion protein was not assayed in cell cultured supernatants by ELISA.

CONCLUSIONSTransfection of HepG2 cells with a chimeric HBsAg-HSP70 construct leads to express fusion protein, but it does not secrete into cell cultured supernatants.

Bacterial Proteins ; Enzyme-Linked Immunosorbent Assay ; HSP70 Heat-Shock Proteins ; genetics ; Hepatitis B Surface Antigens ; genetics ; Hepatitis B Vaccines ; immunology ; Humans ; Immunohistochemistry ; Plasmids ; Recombinant Fusion Proteins ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Vaccines, DNA ; immunology

OBJECTIVETo study the protective effect of intensive insulin treatment on the myocardium of severely scalded rats, and to primarily explore its mechanism.

METHODSEighteen SD rats were divided into three groups, with 6 rats in each group. The rats in burn and intensive insulin group were inflicted with 30% TBSA full-thickness injury on the back. Isotonic saline containing 0.12 U/ml insulin solution, and 100 g/L glucose solution were infused into the rats in the intensive insulin group to keep plasma glucose at the level of 4.0 - 6.6 mmol/L (the total fluid amount was 2 ml x kg(-1) x 8h(-1)). In sham burn group,fluid was given according to physiological demand. The same amount of isotonic saline was infused into the rats in burn group. The venous blood was obtained for the detection of plasma glucose contents, and the left ventricular systolic pressure (LVSP) and left ventricular end-diastolic pressure (LVEDP) were recorded via aortic ventricle cannula before scald and at 1, 2, 3, 4, 5, 6 post-scald hours (PSH). The tissue of the left ventricle was harvested at 6 PSH for the detection of troponin T expression in myocardiocytes.

RESULTSPlasma glucose level was increased to (7.6 +/- 1.7) mmol/L - (8.4 +/- 4.7) mmol/L in burn group during 1-6 PSH, which was significantly higher than that in intensive insulin group (4.5 +/- 0.9) mmol/L - (5.2 +/- 1.3) mmol/L, P < 0.01). Compared with the intensive insulin group, LVSP was markedly decreased in the burn group (60 +/- 11 mm Hg vs 72 +/- 8 mm Hg, P < 0.05) at 1 PSH,whereas LVEDP was increased significantly (21.3 +/- 11.3 mmHg vs 11.7 +/- 5.2 mmHg, P < 0.05). Intensive insulin treatment could significantly inhibit the loss of troponin T protein in myofilaments of myocardium.

CONCLUSIONIntensive insulin treatment possesses a protective effect on myocardia function after severe burns, and it may be related to its preventive effect on the loss of contractile protein in cardiocytes.

Animals ; Blood Glucose ; metabolism ; Burns ; drug therapy ; metabolism ; Insulin ; administration & dosage ; therapeutic use ; Male ; Myocardial Contraction ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley ; Troponin T ; metabolism