Identification of plasma SAA2 as a candidate biomarker for the detection and surveillance of non-small cell lung cancer.

This study aimed to measure the expression of SAA2 in plasma and to assess its diagnostic efficacy as a biomarker for non-small cell lung cancer (NSCLC). The gene expression of SAA2 in NSCLC was analyzed based on a database. Then, SAA2 expression was detected by immunohistochemistry in lung tissue and by enzyme-linked immunosorbent assay in 90 patients with NSCLC and 61 normal controls.

CpG oligodeoxynucleotides enhance chemosensitivity of 5-fluorouracil in HepG2 human hepatoma cells via downregulation of the antiapoptotic factors survivin and livin.

Background: Recent studies indicated that a synthetic oligonucleotide containing un-methylated CpG oligodeoxynucleotides (CpG-ODN) has a potential function for cancer therapy. In this study, we evaluated the chemosensitizing effects of CpG-ODN in 5-fluorouracil (5-FU)-treated HepG2 human hepatoma cells.

Methods: Cell viability assay were utilized to evaluate the direct cytotoxicity of CpG-ODN in the presence or absence of 5-FU in HepG2 cells, and apoptosis as well as cell-cycle was examined by flow cytometry analysis.

Curcumin induces FasL-related apoptosis through p38 activation in human hepatocellular carcinoma Huh7 cells.

Aim: The aim of this study is to explore the underlying molecular mechanism of curcumin-induced apoptosis in human hepatocellular carcinoma (HCC) Huh7 cells.

Main Methods: Fas and FasL mRNA expression was analyzed by reverse transcription PCR. Western blot was applied to detect the protein expression of Bcl-2 family members, MAPK family members, c-Jun, c-Fos, ATF-2, caspase-3, PARP, TNF receptor family members and the respective ligands.

Improvement of lipid profile and reduction of body weight by shan he jian fei granules in high fat diet-induced obese rats.

Background: The goal was to study lipid profiles (TG, TC, LDL, HDL), effects on serum leptin, and fat tissue adiponectin, and resistin as well as body weight effects of Shan He Jian Fei Granules (SHJFG) in rats on a high fat diet.

Methods: Rats were randomly divided into five groups: normal control group fed with normal fat diet, rats on high fat diet receiving low dosage, middle dosage, high dosage of Shan He Jian Fei Granules (SHJFG) as well as a high fat diet group receiving placebo. Rats were treated for 8 weeks.

Promotion of immunity of mice to Pasteurella multocida and hog cholera vaccine by pig interleukin-6 gene and CpG motifs.

A novel oligodeoxynuleotides containing 11 CpG motifs was synthesized and inserted into the VR1020 plasmid containing pig interleukin-6 (IL-6) gene (VPIL6) to construct recombinant plasmid, VPIL6C. The chitosan nanoparticles (CNP) were prepared by ionic cross linkage to entrap the VPIL6C (VPIL6C-CNP), VPIL6 (VPIL6-CNP) and CpG (CpG-CNP). 42-Day old female mice were divided into four groups and intramuscularly injected respectively with 6 pmol VPIL6C-CNP, VPIL6-CNP, CpG-CNP and VR1020-CNP along with the bivalent vaccines against the Pasteurellosis and hog cholera.

Expression of recombination-activating genes and T cell receptor gene recombination in the human T cell leukemia cell line.

Background: Recent studies have suggested that mature T cells can change their specificity through reexpression of recombination-activating genes (RAG) and RAG-mediated V(D)J recombination. This process is named receptor revision and has been observed in mature peripheral T cells from transgenic mice and human donors. However, whether thebreceptor revision in mature T cells is a random or orientated process remains poorly understood.

Quantification of antibody (IgY) titers in hen eggs following immunization and their use in detecting cell surface molecules on nitrocellulose membranes.

HLA-A*0201 alpha chain and beta2m were expressed from a prokaryotic system, and after refolding and purification, the alpha chain and beta2m were used to immunize eight laying hens. The titer of egg yolk antibody against alpha chain increased from 10(2) to 10(5.3) The titer of egg yolk antibody against beta2m increased from 10(1) to 10(4.

Improved preparation of class I HLA tetramers and their use in detecting CMV-specific CTL.

Different methods were used to prepare HLA tetramers and the yields of each method were compared. Our results indicate that preliminary refolding of the heavy chain (Hc) and light chain (beta 2m) yields more monomer than the typical conventional method with urea-solubilized Hc and beta 2m. We then used the corresponding tetramers to detect cytomegalovirus (CMV)-specific cytotoxic T lymphocytes (CTL).

Analysis of the CDR3 length of TCR alphabeta T cells in the peripheral blood of patients with chronic hepatitis B.

To clarify the characteristics of TCR alphabeta T cells in the peripheral blood of patients with chronic hepatitis B (CHB), we investigated the patterns of Complementarity Determining Region3 (CDR3) length distribution for all 24 TCR BV gene families and 32 TCR AV gene family in the peripheral blood lymphocytes of two patients with CHB and two healthy controls by immunoscope spectratyping technique. We found that the profiles of CDR3 length distribution for all TCR AV and TCR BV family showed Gaussian distribution (the polyclonal TCR alphabeta T cells) in healthy controls, however, the restricted usage of TCR BV and TCR AV family (the oligoclonal TCR alphabeta T cells) has been monitored in two CHB patients, furthermore, the oligoclonal TCR alphabeta T cells showed the different CDR3 sequences and length, it might be correlated to the different epitope of hepatitis B virus (HBV) or the different HLA type of the patients. Detailed analysis of the CDR3 length of TCR alphabeta T cells might be interesting to light on the further study of the individualized immunotherapy of CHB and the further research of the new T lymphocyte epitope vaccine of HBV.

Efficient production, purification, and application of egg yolk antibodies against human HLA-A*0201 heavy chain and light chain (beta2m).

The importance of eggs as a source of specific antibodies is well recognized. Egg yolk contains 8--20mg immunoglobulins (IgY) per milliliter. However, the major problem in separating IgY is to remove the high concentrations of lipids in egg yolk.

Effect of transgenic expression of porcine interleukin-6 gene and CpG sequences on immune responses of newborn piglets inoculated with Pseudorabies attenuated vaccine.

Porcine interleukin-6 gene and CpG sequences were used as immunoadjuvants to enhance the immune responses of newborn piglets to Pseudorabies attenuated vaccine (PAV). The titer of specific antibodies to PAV, the proliferation of lymphocytes and induced IL-2 activities were all examined to identify the immune response of the piglets. The results showed that the immune responses with CpG ODN and porcine interleukin-6 gene were significantly stronger than routine immunities.

[High cell density fed-batch culture of E.coli expressing rhVEGF121].

Objective: To investigate the optimal high cell density fermentation conditions of recombinant E.coli BL21/pET- 24a/hVEGF(121) expressing recombinant human vascular endothelial growth factor (rhVEGF(121)).

Methods: The effects of the composition of the fermentation medium, induction time and fed-batch carbon sources on the expression level of rhVEGF(121) and cell output were analyzed.

The protein X4 of severe acute respiratory syndrome-associated coronavirus is expressed on both virus-infected cells and lung tissue of severe acute respiratory syndrome patients and inhibits growth of Balb/c 3T3 cell line.

Background: The genome of the severe acute respiratory syndrome-associated coronavirus (SARS-CoV) includes sequences encoding the putative protein X4 (ORF8, ORF7a), consisting of 122 amino acids. The deduced sequence contains a probable cleaved signal peptide sequence and a C-terminal transmembrane helix, indicating that protein X4 is likely to be a type I membrane protein. This study was conducted to demonstrate whether the protein X4 was expressed and its essential function in the process of SARS-CoV infection.

Immunogenicity of SARS inactivated vaccine in BALB/c mice.

Severe acute respiratory syndrome (SARS) is a serious infectious threat to public health. To create a novel trial vaccine and evaluate its potency, we attempted to generate a SARS inactivated vaccine using SARS coronavirus (SARS-CoV) strain F69 treated with formaldehyde and mixed with Al(OH)3. Three doses of the vaccine were used to challenge three groups of BALB/c mice.

[Preparation and preliminary characterization of soluble HLA-A2-peptide complex].

Aim: To express HLA-A2 heavy chain (HC) and light chain beta(2m) in bacteria and to prepare soluble HLA-A2-peptide complex.

Methods: HC and beta(2m) expressing engineering bacteria was induced for 5 h with 0.5 mmol/L IPTG.

[Preparation and identification of egg yolk antibodies against HLA-A* 0201 heavy chain].

Objective: To prepare a high-titer specific and efficient egg yolk immunoglobulin (IgY).

Methods: The antigen of HLA-A* 0201 heavy chain was used to immunize the hens together with Freud's adjuvant and the eggs these hens laid were collected. IgY was purified from the egg yolk by water extraction, salt precipitation with ammonium sulfate and dialysis.