Prenatal ultrasonographic manifestations and genetic analysis of eight fetuses with 16p11.2 microdeletions / 中华医学遗传学杂志

OBJECTIVE@#To analyze the intrauterine phenotype and genotype of eight fetuses carrying a 16p11.2 microdeletion.@*METHODS@#5100 fetuses undergoing routine prenatal diagnosis were subjected to single nucleotide polymorphism-based microarray (SNP-array) analysis. Fetuses harboring a 16p11.2 microdeletion were analyzed for their ultrasonographic characteristics.@*RESULTS@#Eight fetuses were found to harbor a microdeletion in the 16p11.2 region. Among these, six had a typical 500-600 kb deletion, while the remaining two had an atypical 220 kb deletion at the distal part of 16p11.2. Four fetuses showed vertebral malformations, two had mild left ventriculomegaly, one had hydrocephalus, and one had pulmonary valve stenosis with regurgitation. The parents of five fetuses have accepted pedigree verification, and the results confirmed that the 16p11.2 microdeletions carried by fetuses all had a de novo origin.@*CONCLUSION@#The intrauterine phenotypes of fetuses carrying a 16p11.2 microdeletion may be variable, and the deletion can be effectively detected with the SNP-array assay.

Prenatal ultrasonic characteristics and genetic analysis of fetuses with chromosome 22q11 microdeletion syndrome / 中华医学遗传学杂志

OBJECTIVE@#To analyze the prenatal ultrasonic characteristics and genetic features of 14 fetuses with chromosome 22q11 microdeletion syndrome (22q11DS).@*METHODS@#4989 fetuses were analyzed by using single nucleotide polymorphism array (SNP array) in the Fujian Maternal and Child Health Hospital from November 2016 to November 2019.@*RESULTS@#SNP array showed that 11 fetuses had classic 3 Mb microdeletion in 22q11 region, one fetus had 2.0 Mb microdeletion, and two fetuses had 1.0 Mb microdeletion. The 1.0 Mb microdeletion in 22q11 region contains SNAP29 and CRKL genes, which may increase the risk of congenital renal malformation and cardiovascular malformation.@*CONCLUSION@#Prenatal ultrasonic characteristics of fetuses with 22q11 microdeletion syndrome vary, and SNP array is a powerful tool to diagnose such diseases, which can provide accurate genetic diagnosis and enable prenatal diagnosis.

Prenatal diagnosis and clinical analysis of two fetuses with Cat-eye syndrome / 中华医学遗传学杂志

OBJECTIVE@#To determine the origin of supernumerary small marker chromosomes (sSMCs) carried by two fetuses.@*METHODS@#Single nucleotide polymorphism array (SNP-array) and fluorescence in situ hybridization (FISH) analysis were carried out on cells cultured from the amniotic fluid samples.@*RESULTS@#SNP-array analysis showed both fetuses to be arr[hg19]22q11.1q11.21(16 888 899-18 649 190)×4, with a duplicated 1.7 Mb region (16 888 899-18 649 190) leading to partial tetrasomy of 22q11.1-22q11.21. FISH confirmed that both fetuses were 47,XN,+mar.ish idic(22)(q11.2) (RP11-958H20 ++),which suggested a diagnosis of Cat-eye syndrome (CES). The appearance of abortuses were consistent with the diagnosis of CES.@*CONCLUSION@#Two fetuses with CES were diagnosed by genetic testing. The latter has provided a basis for genetic counseling.

Prenatal diagnosis of 22q11 microdeletion syndrome / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To establish a method for the prenatal diagnosis of 22q11 microdeletion syndrome.</p><p><b>METHODS</b>BACs-on-Beads (BoBs) and fluorescence in situ hybridization (FISH) were performed on a fetus for whom amniotic chromosomal culturing has failed and a pair of twin fetuses suspected for 22q11 deletion syndrome.</p><p><b>RESULTS</b>22q11 microdeletion was detected in all 3 fetuses by prenatal BoBs as well as FISH, with only one red signal detected at the DiGeorge/VCFS N25 site and two green signals on the 22q13.3 ARSA site.</p><p><b>CONCLUSION</b>The combination of prenatal BoBs and FISH can provide a method for the prenatal diagnosis of 22q11 microdeletion.</p>

Mechanism of priming cytotoxic T cell response and strategy for enhancing DNA vaccine potency in DNA immunization / 生物医学工程学杂志

DNA vaccination that can induce both cellular and humoral immune response has become an attractive immunization strategy against cancer and infectious disease. Elucidation of the precise mechanisms of immune priming will be important in the development of effective DNA vaccines. In this review, we illustrate possible mechanisms in priming cytotoxic T cell response involving the intracellular degradation, processing and presentation of encoded antigen. We also discuss the roles of costimulatory molecules expressed on antigen-presenting cells (APCs) in inducing optimal CTL activity. Hence, a rational strategy for increasing DNA potency would be to facilitate these pathways. Additionally, we focus on recent strategies including rapid degradation of ubiquitin-antigen fusion proteins, direct targeting to APCs for increased DNA uptake, direct routing an antigen into the MHC class I and II processing and presentation pathways, and increasing the immunogenicity of encoded antigen. All of these approaches have resulted in increased potency of DNA vaccines.

New type recombinant antibody fragment scFv multimer and cancer targeting / 生物医学工程学杂志

New type recombinant antibody single chain variable fragment (scFv) is formed by the joined VH and VL domains of immunoglobulin with the used of a polypeptide linker that is at least 12 residues in length. scFv is the smallest functional unit of antibody and has shown a fine prospect for the radioimmunoscintigraphy of cancer because of its special characteristics including increased tumour penetration and fast clearance rates compared with parent Ig. A scFv molecule with a linker of 3-12 residues cannot fold into a functional Fv domain and instead associates with a second scFv molecule to form a bivalent dimer (Diabody). Reducing the linker length below three residues can force scFv association into trimers (Triabody) or tetramers (Tetrabody) depending on linker length, composition and V-domain orientation. This review describes linker length and V-domain orientation of scFv, expression and stability of scFv multimers, size of scFv multimers and its effect on in vivo pharmacokinetics, flexibility and avidity of scFv multimers, in vitro application of multimeric murine scFv, multispecific scFv multimers and cancer targeting.

Generation of CD14+ dendritic cells in vitro with GM-CSF and IL-4 / 生物医学工程学杂志

This study was conducted to get high quality and sufficient numbers of mature dendritic cells from healthy donor peripheral blood. The experiment began on culturing of plastic-adherent monocytes isolated from healthy donor peripheral blood with granulocyte-monocyte clony-stimulating factor (GM-CSF 150 ng/ml) and interleukin 4 (IL-4 800 U/ml) without fresh medium feeding and cytokines for 7 days. After 7 days, CD14+ monocytes not only differentiated into high purity DC but also expressed HLA-I and HLA-II molecules, costimulating molecules, adherent molecules and its progenitor marker CD14 molecule highly. These cells displayed all phenotypic and morphologic characteristics of mature dendritic cells and were most potent stimulatory cells in allogeneic mixed leukocyte reactions. The endocytosis ability of these DCs peaked at the third day in culture and decreased remarkably afterwards. These results provide evidence for the first time that CD14+ monocytes differentiated in vitro from peripheral blood monocytes exhibit dendritic cells characteristics and still express its progenitor marker CD14 molecules highly. The results of this experiments may facilitate further studies of CD14+ DC and its clinical applications.

Immune responses of dendritic cells after loaded with cytotoxicity T lymphocyte epitope based peptide of human alpha-fetoprotein (hAFP) / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the immune responses of lymphocytes after activated by dendritic cells (DCs) loaded with cytotoxicity T lymphocyte (CTL) epitope based peptide of human alpha-fetoprotein (hAFP, 218-226 LLNQHACAV).</p><p><b>METHODS</b>Get high purity DCs by cultured plastic-adherent monocytes isolated from healthy donor of HLA-A2(+) peripheral blood with granulocyte-monocyte colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) for 7 days. Stimulate self-lymphocytes with DCs that loaded with CTL epitope based peptide of hAFP under the culture medium contains interleukin-2 (IL-2) for 7 days. Analyse IL-12 and TNF in culture medium and also the specific lysis activity of lymphocytes against four strains of primary hepatocellular carcinoma cells.</p><p><b>RESULTS</b>After stimulated by DC loaded with CTL epitope based peptide derived from hAFP, lymphocytes appeared a good characteristics and the culture medium of activated lymphocytes contained a high level Th1 type cytokines of IL-12 and TNF. Activated lymphocytes not only specifically lysed HLA-A2(+) HepG2 line but also had the cytotoxicity against other three primary hepatocellular carcinoma cell lines and T2 target cell loaded with peptide of hAFP.</p><p><b>CONCLUSIONS</b>The results of this research supply the basic materials for the DC based vaccine with HLA-A2 restricted peptide epitope derived from hAFP against AFP positive primary hepatocellular carcinoma.</p>

Polylatic acid microspheres-induced CTL cytotoxicity against hepatocellular carcinoma cell lines / 中国肿瘤生物治疗杂志

Objective: To investigate the in vitro efficiency of two kinds of polylatic acid (PLA) immunomicrospheres: M1(hAFP158~166)and M2(hAFP218~226) in specific inducement of CTL and the cytotoxicity of CTL against hepatocellular carcinoma cells. Methods: Pripheral blood monocytes (PBMC) of HLA-A2+ healthy volunteers stimulated in vitro by peptide-loading microspheres were taken as effectors. The experiments were divided into three groups: control group, hAFP+ tumor cells group, and hAFP- tumor cells group. The specific cytolysis of target cells was assessed by 51Cr-release assay. Results: Both M1 and M2 induced proliferation of HLA-A2+ PBMC, forming visible clones. The effector cells induced by M1 and M2 both had an cytolysis rate over 75% for hAFP+ tumor cells, which was significantly higher than that for hAFP- tumor cells (P0.05). Conclusion: The two kinds of microspheres can both induce specific CTL in vitro, which can effectively kill hAFP+ tumor cells.

Study on the Immune regulation of anti-idiotypic antibodies against anti-HBs in HBV infection——一、The production of anti idiotypic antibodies against anti HBs / 中国免疫学杂志

The anti-idiotypic antibodies Specific for anti-HBs were prepared by Immunizing New Zealand white rabbits with anti-HBs according to the method of Kennedy. The serum was first purified through IgG of normal human-Sepharose 4B affinity Column.The anti-idiotypic Specificity of the antibodies obtained was identified with ELISA or RIA inhibition test.The resultshowed that the anti-idiotypic antibodies were directed at the idiotypic determinants on anti-HBs,and no cross reaction between anti-idiotypic antibodies and normal human IgG.