Objective To develop an effective and broad immune protective H5N1 vaccine.Methods We first developed two recombinant vaccinia ( Tiantan strain) virus ( rTTV ) based H5N1 vaccines, which consisted of bicistron expressing the hemagglutinin(HA) and matrix protein 2(M2), or bicistron expressing the neuraminidase(NA) and matrix protein 1 (M1). The expression of H5N1 protein in rTTVs was confirmed. We immunized the BALB/c mice twice with two kind of dose ( 104 PFU, 107 PFU)using different combination. Subsequently, we assessed the humoral and cellular immune response in vaccinated mice. Results Our data showed that rTTV-based H5N1 vaccine induced rapidly robust HA- and NAspecific antibody level and IFN-γ secreting form cell(SFC) with either single dose of 107 PFU or twice dose of 104 PFU or 107 PFU. We also detected significant neutralizing antibody and matrix-specific immune response. In addition, we found that immunization with two kind of rTTV-based H5N1 vaccines induced much high level of M2-specific antibody than that with single of rTTV-based H5N1 vaccine. Conclusion rTTVbased H5N1 vaccines in this study elicited board array of immunity and our study offers a promising alternative H5N1 vaccine candidates with favorable potential to prevent various H5N1 pandemic.

Objective To characterize the immunogenicity in gene immunization of the conserved regions of hepatitis C virus(HCV) based on different delivery strategies. Methods We first constructed a novel DNA vaccine encoding a fusion gene(from partial NS3 and Core) of HCV. Then we compared different protocols based on naked DNA injection twice or DNA injection with gene electrotransfer(GET) in BALB/c mice. The immune response was measured by antibody ELISA and by IFN-gamma ELISPOT. Results Our data showed that a protocol based on intradermally injection of DNA with optimal GET induced the strongest humoral and cellular immunity, and DNA with GET induced a substantially higher anti-NS3/Core T cell re-spoase than naked DNA injection. Conclusion Our data suggest that DNA vaccines encoding NS3/Core fu-sion protein of HCV immunized by the present strategy could merit further study in the context of future prophylactic and therapeutic HCV T cell based vaccines.

Recombinant adeno-associated viruses (rAAV) vectors have been shown to mediate long-term transgene expression in mice and nonhuman primates. We have adapted viral vector system based on two rAAV vectors, namely rAAV1 and rAAV2. We have generated rAAV vectors expressing the envelope glycoprotein (E1 and E2) derived from Chinese HCV patient (genotype 1b) and used these to immunize BALB/c mice. We detected the total antibody titer by IFA and neutralizing antibody (nAb) using in vitro HCV neutralizing assays based on HCV pseudotyped particles. Furthermore, IFN-gamma ELISpot assay was used to assess the T cellular response against HCV at 12 weeks after rAAV1-E1E2 immunization. We also analyzed HCV envelope glycoprotein expression in muscle of rAAV1-E1E2 immunized mice. Our data showed: (i) rAAV1 directed long-term expression of HCV genes in mice; (ii) immunized intramuscularly with a single dose of rAAV elicited durable and effective immune responses in mice; and (iii) Moreover, rAAV1-E1E2 induced higher total antibody and nAb titers than rAAV2-E1E2 did. These data suggest that rAAV1 vectors could stimulate robust, durable, and effective immune responses against HCV.
Animals ; Antibodies, Viral ; blood ; Dependovirus ; genetics ; metabolism ; Female ; Genetic Vectors ; genetics ; metabolism ; Hepacivirus ; genetics ; immunology ; Hepatitis C ; immunology ; virology ; Humans ; Mice ; Mice, Inbred BALB C ; Vaccines, DNA ; administration & dosage ; genetics ; immunology ; Viral Envelope Proteins ; administration & dosage ; genetics ; immunology ; Viral Vaccines ; administration & dosage ; genetics ; immunology

To compare different adjuvant formulation and explore the impact of Calcineurin B subunit(CnB) as adjuvant with a novel HBV protein particle (HBSS1) vaccine in mice, female C57BL/6 mice were immunized HBSS1 with Al(OH)3 only, or a normal dose (5 μg) CnB only, or (CnB+ Al(OH)3) mixture as the adjuvant. All immunized groups were primed twice at 4-week intervals; followed by boosting with recombinant adenoviral based HBV vaccine(rAdSS1) at 10-week intervals. We detected the antigen specific humoral response in mice, including total IgG antibody and IgG subtyping. Then, we characterized the specific cell-mediated immune (CMI) response by detection of γ-interferon secreting splenocytes after stimulaton with S or PreS1 peptide pools. No enhancement of immunity was found among the mice with 5 μg of CnB alone or combined with Al(OH), adjuvanted vaccine,which could not induce higher level of anti-PreS1 and anti-S antibodies and CMI than that of HBSS1 alone or Al(OH)3 adjuvanted vaccines. We concluded that CnB is not an effective adjuvant for a novel HBV subunit vaccine.
Adjuvants, Immunologic ; pharmacology ; Animals ; Calcineurin ; pharmacology ; Female ; Hepatitis B Vaccines ; immunology ; Mice ; Mice, Inbred C57BL ; Protein Subunits

This study was purposed to investigate the change of Th cell subsets in the patients with acute graft-versus host disease (aGVHD) and to explore its role in the pathogenesis of aGVHD after allogeneic hematopoietic stem cell transplantation (allo-HSCT). 23 patients underwent allo-HSCT were selected for analysis. The aGVHD in patients was diagnosed according to clinical features, and was confirmed by skin biopsy in some patients. The peripheral blood from 23 patients was collected before and after allo-HSCT. The quantitative chenges of Th1 and Th2 cells in peripheral blood samples were detected by using flow cytometry. The results showed that out of 23 patieats the aGVHD occured in 8 patients including 1 case of grade I, 2 case of grade II, 3 cases of grade III; no aGVHD occured in 15 patients. The flow cytometry analysis revealed that the amount of Th1 cells in patients with aGVHD was much higher than that in patients without aGVHD (p < 0.01), the IFN-gamma expression of Th1 cells in patients with aGVHD of grad II - III significantly was higher than that in patients without aGVHD (p < 0.01), meanwhie the IL-4 expression of Th2 cells in patients with aGVHD of grade II - III was significantly lower than that in patients without aGVHD (p < 0.05). Dynamical detection indicated that the Th1 obviously increased before occurrence of aGVHD and before treatment of aGVHD, while the Th1 cells obviously decreased after treatment of aGVHD. The Th1 cells not changed significantly in patients without aGVHD before and after allo-HSCT. It is concluded that Th1 cells obviously increase in patients with aGVHD, this increase is related to aGVHD pathogenesis. Detecting the changes of Th cell subsets in the early stage after allo-HSCT may be contributed to early diagnosis and therapy of aGVHD.
Graft vs Host Disease ; blood ; immunology ; Hematologic Neoplasms ; immunology ; therapy ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Th1 Cells ; immunology ; pathology

Objective To find out the differences of dietary patterns among freshmen coming from urban and rural areas that might have influenced their bone mineral density and body mass index (BMI).Methods With stratified random sampling method,dietary patterns and their bone mineral density,BMI of 1319 freshmen were studied.Results (1) The ratios of urban freshunen who chose "western food" pattern ( x2=31.548,P=0.000; x2=13.068,P=0.001 ),"animal food" pattern ( x2=8.279,P=0.016; x2=41.137,P=0.000) or "calcium food" pattern (x2=37.254,P=0.000; x2=15.651,P=0.000) were higher than that of rural freshmen,and the ratios of rural freshmen who chose "Chinese traditional" pattern (x2=36.194,P=0.000; x2=25.936,P=0.000) were higher than that of urban freshmen.(2) The average height,weight,BMI,speed of sound (SOS) of male freshmen from rural areas were lower than that from the city and the differences were statistically significant (P＜0.05).Among those female freshmen,only height and weight were significantly different (P＜0.05).(3) In both rural and urban frestunen,the factor scores of "westem food" pattern had a positive correlation with BMI,with the correlation coefficients as 0.187,0.192,0.551,0.465 (P＜0.001).The factor scores of "calcium food" pattern were positively related to bone mineral density (SOS values)with correlation coefficients as 0.680,0.342,0.841,0.786,P＜ 0.001 respectively.The factor scores on "Chinese traditional" pattern were negatively correlated with BMI,with correlation coefficients as -0.223,-0.093 (P＜0.05) which were positively related to bone mineral density (SOS values) in both rural and urban male freshmen,with correlation coefficients as 0.905,0.711 (P＜0.001).Conclusion Different dietary patterns chosen by urban and rural freshmen had a significant impact on both bone mineral density and BMI.

To develop a new Hepatitis C virus (HCV) DNA vaccine ,We explored strategies for optimizing the immunogenicity of HCV DNA vaccine in fusion with dendritic cell-targeting molecules (scDEC, a single-chain antibody against the murine DC cell surface molecule DEC205). We constructed the DNA vaccine plasmids expressing the HCV non-structural protein NS3 alone or in combination with DEC205 as a fusion protein and identified the expression of the molecules of interest by transient transfection of 293 cells with the resultant DNA vaccine plasmids. Then BALB/C mice were immunized with these plasmids by the injection in combination with electroporation. The NS3-specific IgG antibody(ELISA) and cellular immunity (IFN-gamma ELISPOT) were analyzed post twice immunization. Our results showed that: the single-chain antibody against DEC205 fused with vaccine antigen NS3 significantly enhanced the immunogenicity of new HCV DNA vaccine, the intradermal injection in combination with electroporation using caliper electrodes resulted in most robust NS3-specific antibody and T cell immune response. In conclusion,immune response for the HCV NS3 protein-encoding DNA vaccine was enhanced significantly when targeting antigen NS3 to DCs by scDEC. The present strategy could merit further study in the context of other prophylactic and therapeutic DNA based vaccines.
Animals ; Cell Line ; Dendritic Cells ; immunology ; virology ; Enzyme-Linked Immunospot Assay ; Female ; HIV Antibodies ; immunology ; Hepacivirus ; genetics ; immunology ; Hepatitis C ; immunology ; prevention & control ; virology ; Humans ; Mice ; Mice, Inbred BALB C ; Vaccines, DNA ; administration & dosage ; genetics ; immunology ; Viral Nonstructural Proteins ; administration & dosage ; genetics ; immunology

This study aims to develop inexpensive and effective experimental vaccines against highly pathogenic H5N1 Avian Influenza (HPAI) virus and to optimize their immunization programs. To this end, we first synthesized the codon-optimized hemagglutinin gene (HAop) and neuraminidase gene (NAop), both of which were derived from a H5N1 virus (Anhui strain), and constructed successfully the DNA vaccines containing a single cistronic construct (HAwt, HAop, or NAop) or a bicistronic construct (HAop/M2 or NAop/M1) of H5N1 influenza virus origin. Their expression was confirmed by indirect immunofluorescent assay (IFA) and Western blotting. Then twice vaccination of mice with the DNA vaccines by injection intramuscularly or in vivo electroporation (EP) via two different routes was evaluated and analyzed by hemagglutination inhibition (HI) assay, NA-specific antibody detection, micro-neutralizing antibody test and IFN-gamma ELISpot assay. Our results showed that the DNA vaccines with coden-optimized HAop and NAop constructs could quickly elicit a strong immune response by in vivo EP, especially the cellular immune response against HA and NA; the in vivo EP via intradermal route induced stronger humoral immune responses than those via intramuscular route. Our findings will pave a way for further development of novel DNA-based H5N1 vaccine and for the optimization of the immunization programs of DNA vaccine.
Animals ; Antigens, Viral ; immunology ; Codon ; genetics ; Electroporation ; Female ; Humans ; Influenza A Virus, H5N1 Subtype ; immunology ; Mice ; Mice, Inbred BALB C ; Vaccination ; methods ; Vaccines, DNA ; genetics ; immunology ; metabolism ; Viral Structural Proteins ; immunology

OBJECTIVETo provide genetic consulting and prenatal diagnosis for two families with congenital factor V deficiency based on the known mutations of factor V gene (G16088C and G69969T).

METHODSChorionic DNA was obtained at 12 weeks of gestation and analyzed to exclude maternal cell contamination through microsatellite DNA analysis. It was then amplified with PCR and sequenced to determine the presence of mutations in exons 3 and 23. Factor V activity of the blood was assayed at 22 weeks of gestation and 6 months after birth.

RESULTSThe fetus in case 1 was found to be a heterozygous carrier of the G16088C mutation, for whom factor V activity of the cord blood and peripheral blood were 15% and 53%, respectively. Fetus 2 did not carry the familiar G69969T mutation, for whom the factor V activity of cord blood and peripheral blood has measured 32% and 93%, respectively. Follow-up studies demonstrated that the two infants were both in good health without a tendency for bleeding.

CONCLUSIONIn both cases, the genotypes were consistent with the phenotypes. This is the first report of prenatal diagnosis of congenital factor V deficiency.

Adult ; Base Sequence ; Child ; Factor V ; genetics ; metabolism ; Factor V Deficiency ; diagnosis ; genetics ; Female ; Humans ; Male ; Microsatellite Repeats ; Mutation ; Pregnancy ; Prenatal Diagnosis

Material properties and hygroscopicity were determined. Principal component analysis and partial least squares regression were applied to evaluate relationships between material properties and hygroscopicity of Chinese medicine raw materials. The results showed that hygroscopicity was correlated with water content, particle size distribution, water soluble characteristic and cohesion. Balanced moisture content was positively correlated with water content, particle size distribution, water soluble characteristic and cohesion. Moisture absorption velocity was negatively correlated with particle size distribution, protruding degree and positively correlated with water soluble characteristic and cohesion. Moisture absorption acceleration was positively correlated with water content, particle size distribution and negatively correlated with water soluble characteristic and cohesion. Hygroscopicity of Chinese medicine raw materials is interpreted in terms of physics.
Absorption ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Least-Squares Analysis ; Multivariate Analysis ; Particle Size ; Plants, Medicinal ; chemistry ; Principal Component Analysis ; Solubility ; Water ; analysis ; Wettability