1.CpG-ODN plus alum as a composite adiuvant to enhance the immunogenicity of influenza split virion vaccine
Xing LUO ; Ying LIU ; Chunting YANG ; Minqing GONG ; Jun ZHOU ; Sanke YU ; Honglin XU
Chinese Journal of Microbiology and Immunology 2011;31(10):942-947
Objective To evaluate the immuno-potentiating effects of CpG-ODN plus alum as a composite adjuvant on influenza split virion vaccine.Methods BALB/c mice were immunized with various amounts of 2009 H1N1 influenza split virion vaccine,alone or in combination with CpG-ODN,alum,or both (composite adjuvant).Antigen-specific humoral immune responses were evaluated by ELISA,hemagglutination inhibiting (HI) assay and neutralizing assay.Antigen-specific cellular immune responses were evaluated by ELISPOT assay,intracellular cytokine staining assay and in vivo CTL assay.Results Compared with the control group immunized with antigen alone,a single use of either adjuvant weakly enhanced the humoral immune responses,as indicated by the increase of antigen-specific IgG titers,HI titers and neutralizing titers by 3-6 folds,2-4 folds and 4-8 folds,respectively,after two immunizations.In contrast,the composite adjuvant induced more potent humoral immune responses; the antigen-specific IgG titers,HI titers and neutralizing titers were increased by 23-57 folds,9-20 folds and 16-64 folds,respectively.Consequently,the composite adjuvant achieved antigen-sparing by at least 16 folds.In addition,the composite adjuvant significantly enhanced the antigen-specific cellular immune responses,as revealed by the increase of IFN-γ-secreting CD4+ T cells and the enhancement of CTL activity in immunized mice.Conclusion CpG-ODN plus alum as a composite adjuvant can enhance the immunogenicity of influenza split virion vaccine and achieve the antigen-sparing effect.
2.Role of PI3K∕Akt signaling pathway in propofol?induced invasion of human liver cancer cell line HepG2
Qiaoling ZHOU ; Hongzhen LIU ; Minqing YANG ; Yuanying LIU ; Xiaohong LAI ; Hua LIANG ; Chengxiang YANG ; Miaoning GU
Chinese Journal of Anesthesiology 2018;38(1):110-113
Objective To evaluate the role of phosphatidylinositol 3?kinase(PI3K)∕serine?threo?nine kinase(Akt)signaling pathway in propofol?induced invasion of human liver cancer cell line HepG2. Methods HepG2 cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum in a 5% CO2incubator at 37℃. HepG2 cells at the logarithmic growth phase were divided into 6 groups(n=18 each)using a random number table: control group(group C), propofol group(group P), PI3K∕Akt signaling pathway agonist IGF?1 group(group IGF), PI3K∕Akt signaling pathway inhibitor LY294002 group(group LY), IGF?1 plus propofol group(group IGF+P)and LY294002 plus propofol group (group LY + P). Propofol 120 μg∕ml was added in group P. IGF?1 10 nmol∕L was added in group IGF. LY294002 10 μmol was added in group LY. In group IGF+P, 10 nmol∕L IGF?1 was added, cells were in?cubated for 24 h, and then 120 μg∕ml propofol was added. In group LY+P, 10 μmol LY294002 was add?ed, cells were incubated for 24 h, and then 120 μg∕ml propofol was added. The invasion of cells was measured by Transwell invasion assay at 24 h of incubation. The expression of PI3K and Akt mRNA in cells was determined by real?time polymerase chain reaction. The expression of Akt, PI3K and phosphorylated Akt(p?Akt)was detected by using Western blot. Results Compared with group C, the invasive cell count was significantly reduced, the expression of PI3K protein and mRNA was down?regulated, p?Akt∕Akt ratio was decreased, and the expression of Akt mRNA was down?regulated in P, P+IGF, LY and P+LY groups, and the invasive cell count was significantly increased, the expression of PI3K protein and mRNA was up?regulated, p?Akt∕Akt ratio was increased, and the expression of Akt mRNA was up?regulated in group IGF(P<0.05). Compared with group P, the invasive cell count was significantly increased, the expression of PI3K protein and mRNA was up?regulated, p?Akt∕Akt ratio was increased, and the expres?sion of Akt mRNA was up?regulated in group P+IGF, and the invasive cell count was significantly reduced, the expression of PI3K protein and mRNA was down?regulated, p?Akt∕Akt ratio was decreased, and the ex?pression of Akt mRNA was down?regulated in group P+LY(P<0.05). The invasive cell count was signifi?cantly reduced, the expression of PI3K protein and mRNA was down?regulated, p?Akt∕Akt ratio was de?creased, and the expression of Akt mRNA was down?regulated in group P+IGF as compared with group IGF (P<0.05)and in group P+LY as compared with group LY(P<0.05). Conclusion The mechanism by which propofol inhibits invasion of HepG2 cells is related to inhibiting activation of PI3K∕Akt signaling path?ways.
3.Results of carrier screening and prenatal diagnosis for FMR1 gene in 819 cases.
Jiao LI ; Juan DU ; Qi YANG ; Juanjuan XU ; Meng LI ; Huayu FU ; Minqing LI
Chinese Journal of Medical Genetics 2020;37(10):1104-1107
OBJECTIVE:
To determine the carrier rate of Fragile X mental retardation 1 gene (FMR1) mutants in women with a history of adverse pregnancy or childbirth, and to provide prenatal diagnosis for the carriers.
METHODS:
Peripheral blood samples were collected from women with a history of adverse pregnancy or childbirth, and the FMR1 gene cytosine-guanine-guanine repeat number (CGG)n was determined by triple-repeat primer polymerase chain reaction (TP-PCR) combined with capillary electrophoresis. Prenatal diagnosis was provided for the carriers during pregnancy.
RESULTS:
Among 819 samples, 9 gray zone repeats carriers and 10 premutation carriers were detected, which gave a prevalence of 1 in 91 and 1 in 82, respectively, with a total prevalence of 1 in 43. Prenatal diagnosis was provided during 7 pregnancies for 6 carriers. A female fetus with premutation (n = 30/57) and an affected male fetus with full mutation (n = 336) were detected.
CONCLUSION
FMR1 gene testing in women with a history of adverse pregnancy or childbirth can facilitate genetic counseling and reproductive guidance for carriers of gray zone repeats and premutations. Prenatal diagnosis for carriers of premutation can facilitate reduction of the birth of children with fragile X syndrome.