Serum soluble CD40 is associated with liver injury in patients with chronic hepatitis B.

Soluble cluster of differentiation 40 (sCD40) is proteolytically cleaved from membrane-bound CD40 and binds to CD154, thereby inhibiting CD40-CD154-mediated immune responses. The aim of the present study was to clarify the role of sCD40 in chronic hepatitis B (CHB). The sCD40 levels in sera from 132 patients with CHB and 33 healthy individuals were retrospectively measured.

[Cloning and expressing of tissue inhibitor of metalloproteinases I gene fragment and preparation of monoclonal antibodies against the recombinant protein].

Objective: To prepare the monoclonal antibody (mAb) against tissue inhibitor of metalloproteinases I (TIMP-I) fusion protein.

Methods: TIMP-I gene was amplified from fibrotic human liver tissue by RT-PCR, then ligated with pQE31 to form recombinant plasmid pQE-TIMP-I and transformed into E. coli BL21.

[Analysis of hemagglutination inhibition antibody level in patients with influenza A H1N1].

Objective: To understand the hemagglutination inhibition antibody level in patients with influenza A H1N1.

Methods: Sera from 28 patients with influenza A H1N1 at different time points after illness onset were collected and measured by hemagglutination inhibition assay.

Results: The serum hemagglutination inhibition antibody titers at 1, 5, 15, 22, 37, 49 and 58 days after illness onset were 5.

[Clinical study on hybrid bioartificial liver supporting system for acute on chronic liver failure patients].

Objective: To construct an hybrid bioartificial liver supporting system, and observe its effectiveness and safety on patients with acute on chronic liver failure.

Methods: Hybrid bioartificial liver supporting system (HBALSS) was constructed using bioreactor with HepG2 cells transfected with human augmenter of liver regeneration (hALR) gene. 12 acute on chronic liver failure patients were divided into 2 groups randomly.

[Construction of recombinant plasmid expressing S1 gene of new type of reovirus].

Objective: To construct the recombinant plasmid containing S1 gene of new type of reovirus, and to study the expression of protein sigma1 in Vero cells.

Methods: The recombinant plasmid, named pC-S, was constructed by cloning S1 gene into vector pCAGGS/MCS. Then Vero cells were transfected with pC-S and collected at 24, 48, 72 h post transfection followed by SDS-PAGE and Western-Blot assay.

[Construction and identification of a vector inserted with gene of T7 RNA polymerase].

Objective: To develop a system to rescue virus by intracellular expression of T7 RNA Polymerase.

Methods: The gene of T7 RNA Polymerase was amplified and cloned to VR1012 by molecular biological technology. The expression plasmid VR-1a was then identified.

[Construction and identification of attenuated Salmonella which harboring enterovirus 71 VP1 gene].

Objective: To develop attenuated Salmonella which harboring enterovirus 71 (EV71) VP1 gene.

Methods: The plasmid which expressed VP1 protein of EV71 was constructed by gene recombination. Cellular expression was assessed by Western Blot analysis.

Immunologic changes during Pandemic (H1N1) 2009, China.

We analyzed changes in immunologic values over time for 28 hospitalized patients with pandemic (H1N1) 2009. Levels of interleukin-6, interferon-y, and interleukin-10 increased 1 day after illness onset and then decreased to baseline levels. Levels of virus-specific antibody were undetectable 1 day after illness onset and peaked 36 days later.

[Construction and expression of a coxsackievirus A16 VP1 gene plasmid which delivered by live attenuated Salmonella].

Aim: To develop a coxsackievirus A16 (Cox A16) VP1 gene plasmid which delivered by live attenuated Salmonella.

Methods: The plasmid which expressed VP1 protein of CoxA16 was constructed by gene recombination. Cellular expression was assessed by Western bloten analysis.

[Construction and experimental study on off-line hybrid bioartificial liver supporting system with human liver cell line].

Objective: To construct an off-line hybrid bioartificial liver supporting system with human liver cell line, and study it's effect on the plasma from patients with liver failure.

Methods: We established the bioreactor using Psu-2s (Fresenius) cultured with Hep G2 cell transfected with human augmenter of liver regeneration (hALR) gene, then constructed a hybrid bioartificial liver supporting system, at last using the bioartificial liver support system to purify the plasma treated 2 hours with serum bilirubin absorbent, separated from acute on chronic liver failure patients infected by hepatitis B virus.

Results: Bioreactor was successful constructed.

[Construction and identification of a vector inserted with complete genome of poliovirus strain Sabin I].

Objective: To develop a vector inserted with complete genome of poliovirus strain Sabin I.

Methods: The 3 fragments of the complete genome of Sabin I was amplified and cloned to pEASY-T3 by molecular biological technology. These cloned pEASY-T3 were then digested by Restriction enzymes and ligated to pWSK29 step by step and identified.

[Construction and expression of hepatitis B virus envelope protein combined with core protein with two multiple cloning sites vector].

Objective: To develop a coexpression plasmid which expressing envelope protein and nucleoprotein of hepatitis B virus and know of its expressing efficiency.

Methods: The plasmid coexpressing envelope protein and nucleoprotein of hepatitis B virus under the CMV promoter respectively was constructed by gene recombination. Cellular expression was assessed by ELISA.

[Immune effects of mutated hepatitis B virus precore-core DNA vaccines in mice].

Objective: To observe the immune effect of DNA vaccines encoding mutated HBV pre-c/c gene (VE2,VE4) in mice.

Methods: Three kinds of plasmid VEC(DNA vaccines encoding HBV pre-c/c gene), VE2 and VE4 were injected into the thigh muscles of different group of BALB/c mice.Blood and splenocytes from mice were isolated at 4 weeks after immunization.

[Primry research of separation and culture of adult hepatocytes].

Objective: To explore the separation and culture method of adult hepatocytes.

Methods: The isolated adult hepatocytes were cultivated by RPMI 1640 medium at 37 degrees C in vitro. The characteristics of the growing hepatocytes were observed.

[Level and clinical significance of soluble CD40 in patients with chronic hepatitis B].

Objective: To understand the level and clinical significance of soluble CD40 in patients with chronic hepatitis B.

Methods: Detecting the concentration of sCD40 from 176 cases with chronic hepatitis B by ELISA and analyzing its relationship with different grades of inflammation and necrosis in liver tissue.

Results: sCD40 from patients with chronic hepatitis B was significantly higher than those from healthy.

[A five-year analysis of HBV mutations in a multidrug-resistant patient with chronic hepatitis B].

Objective: To investigate HBV mutations in reverse transcriptase (RT) gene and precore/basal core promoter (PC/BCP) regions in a chronic hepatitis B patient and to analyze the link between the mutations and drug resistance or HBeAg sero-conversion.

Methods: Eighteen serum samples were collected from a chronic hepatitis B patient during his 14 hospitalizations from June 2002 to September 2007. HBV DNA was extracted and nested PCR was employed for amplification of target gene fragments.

[Genome analysis of a newly isolated enterovirus].

Objective: To demonstrate molecular characterization of a newly isolated enterovirus.

Methods: Virus were isolated from patient with unknown-causing disease and tested by reverse transcription-polymerase chain reaction (RT-PCR) and 5'3'RACE (rapid amplification of cDNA ends, RACE), in an attempt to obtain the sequence of this newly isolated enterovirus.

Results: Sequence analysis showed that this newly isolated enterovirus shared 83%-94% nucleotide identity and 91%-100% amino acid identity with enterovirus 89.

[Establishment and evaluation of the diagnostic kit for anti-HIV1/2 antibody and P24 antigen].

Objective: To establish and evaluate an Enzyme Immunoassay diagnostic kit combined with anti-HIV1/2 antibody and P24 antigen for shortening the examination window period of HIV infection in HIV laboratory diagnosis.

Methods: The enzyme-linked reaction plates was coated by anti-HIV P24 monoclonal antibody and HIV 1/2 antigen. Labeling HIV1/2 antigen and anti-HIV P24 polyclonal antibody with horseradish peroxidase, setup an integrated ELISA kit for detecting anti-HIV-1/2 antibody and HIV P24 antigen, and evaluate the specificity and sensitivity of this kit.

[Construction and antigenic evaluation of a recombinant MVA virus-like particle expressing HBV C gene].

Objective: To construct the virus-like parcel expressing hepatitis B virus (HBV) C gene and identify its immunogenicity.

Methods: HBV C gene was cloned into the shuttle vector pSC11, and the resulted plasmid pSC11-C was transfected into modified vaccinia virus Ankara (MVA).

Results: pSC11-C was correctly constructed as verified by sequence analysis and PCR, and the recombinant virus-like parcel possessed good immunogenicity.

[Expression, purification, and characterization of an anti-human RBC ScFv-HIV gp160 fusion protein for hemagglutination-based rapid detection of antibodies to HIV in whole blood].

Objective: To construct and express anti-human RBC and HIVgp160 fusion protein for rapid detection of antibody to HIV.

Methods: The gene of the anti human RBC ScFv and HIV antigen were constructed together into expression vector. The fusion protein was expressed in E.

[Follow up study on viruses associated with SARS among the SARS patients].

Background: To study the existence status of the SARS-CoV, retrovirus, and the poliovirus in the bodies of the patients with SARS and the possible relationship between the three viruses and SARS.

Methods: The clinical specimens of the nasopharyngeal swabs, sputum (or saliva), urine, fecal specimens were collected on three consecutive days from 8 patients with SARS 2 years after the recovery from SARS. SARS-CoV, reovirus and poliovirus RNA was detected by using reverse transcription (RT)-PCR; IgG antibody to the poliovirus type 1 and 3 and the antibody to SARS-CoV were determined using enzyme linked immunosorbent assay (ELISA).

[Identification and typing of human enteroviruses using an RT-PCR assay].

Background: To establish a molecular detection and typing assay for identification and typing of human enteroviruses (HEV) which is suitable for clinical detection and epidemiologic research.

Methods: Using both primers specific for HEV genus and HEV typing primers and reverse transcription polymerase chain reaction (RT-PCR) the authors detected preliminarily HEV by agarose gel electrophoresis and then identified serotype through nucleotide sequence analysis of RT-PCR amplicons. The monospecific antisera neutralization was applied to validate the typing results.

[Preliminary study on determination of specific cellular immunity of patients with hepatitis B].

Background: To evaluate an enzyme linked immunospot (ELISPOT) method for testing the specific cellular immunity of patients with hepatitis B and preliminarily investigate into the difference of cellular immunity in patients with various types of hepatitis B.

Methods: The patients with acute hepatitis B, chronic hepatitis B liver cirrhosis, healthy persons with HBV vaccine immunization, healthy persons with past HBV infection and HBV naive persons were enrolled in this study. Their peripheral blood mononuclear cells were tested by ELISPOT to determine the number of gamma-interferon secreting cells.

[Prokaryotic expression and purification of human immunodeficiency virus p24 antigen].

Objective: To express and purify the HIV p24 gene in E. coli cells, and identify p24 antigen activity.

Methods: The full length gene fragment of HIV p24 was amplified by PCR and inserted into the pRSET vector in order to construct the pRSET-p24 recombined vector.

[Study on the response of specific antibodies against SARS-CoV in patients infected with SARS].

Objective: To study the response of specific antibodies against severe acute respiratory syndrome (SARS)-CoV in patients infected with SARS.

Methods: IgM-capture, indirect and antigen-sandwiched enzyme linked immunosorbent assay (ELISA) were used to detect the SARS-CoV specific IgM, IgG and total antibodies in sera of clinical SARS patients or non-SARS individuals.

Results: The positive rates of IgM, IgG and total antibodies to SARS-CoV in 146 sera of SARS patients collected in different phases of the disease were 61.