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.

Objective To identify the potential risks in the process of nursing care for children with tracheal foreign body removal and improve the skills and the process, to promote the quality of nursing care and the success rate of tracheal foreign body removal. Methods The failure mode and effect analysis (FMEA) were used to analyze the potential risks in children with tracheal foreign body removal, preoperative, intraoperative and postoperative care, and to find out the possible risks in each medical treatment. For pediatric venous puncture technology was not skilled, surgical position placement inappropriate (especially in the case of foreign body removal surgery posture), surgical instruments and equipment management, intraoperative nursing cooperation and observation of the disease, postoperative transfer and other aspects of the transfer of training and improvement. Results The medical risk value (RPN) in the treatment of pediatric tracheal foreign body removal in the process of nursing in the control group was reduced from 1 284.2 to 213.2 of the experimental group. The average value of doctors' satisfaction was increased from 79.33% in the control group to 93.33%in the experimental group, P<0.05, the difference was statistically significant. Conclusions The use of FMEA in the treatment of pediatric tracheal foreign body removal surgery care and medical risk management improve the quality of surgical care and help improve the success rate of tracheal foreign body removal. It also ensures the safety of the children in the operation process and improves the quality of nursing work.

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.

OBJECTIVE:To provide the clinical departments with a reference for promoting rational drug use. METHODS:The inpatient prescriptions given by orthopaedics and burn departments from Jun. to Dec. 2013 were selected randomly. The number of the selected prescriptions by each department accounted for 2% of the total prescriptions given by the corresponding department in that month,including 4 921 cases by orthopaedics department and 1 391 cases by burn department. Statistical analysis of irrational prescriptions was made according to Chinese Pharmacopoeia,New Materia Medica,the package insert, and other relevant references. RESULTS:1 821 prescriptions given by orthopaedics department and 378 by burn department were found to be irrational,accounting for 37.00% and 27.17% respectively. The reasons of irrationality mainly included im-proper compatibility,improper route of administration,contraindication in and use with caution by the elderly and children,re-peated drug use,improper drug combination etc. CONCLUSIONS:The system of prescription review should be strengthened, and clinical staff training system and pharmaceatical knowledge information platform are established to promote rational use of drugs.

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.
Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Dendritic Cells ; cytology ; Granulocyte-Macrophage Colony-Stimulating Factor ; chemistry ; Humans ; Interleukin-4 ; chemistry ; Lipopolysaccharide Receptors ; metabolism ; Monocytes ; cytology

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.
Antibodies, Bispecific ; Immunoglobulin Fragments ; Immunoglobulin Heavy Chains ; chemistry ; Immunoglobulin Light Chains ; chemistry ; Immunoglobulin Variable Region ; Immunotherapy ; Neoplasms ; immunology ; therapy ; Protein Engineering ; Recombinant Fusion Proteins

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.
Animals ; Antigen Presentation ; Antigen-Presenting Cells ; immunology ; Lysosomes ; immunology ; Mice ; T-Lymphocytes, Cytotoxic ; immunology ; Ubiquitin ; physiology ; Vaccines, DNA ; genetics ; immunology

Objective To analyze the urine trace albumin(mALb)/creatinine(Cr) ratio and blood uric acid(UA),and other various metabolic index level in patients with diabetic nephropathy(DN),combined with clinical data such as patients' age,body mass index(BMI),course of diseases,to explore the related mechanism of occurrence and development of DN.Methods 76 DN patients were selected.The microalbuminuria group(urinary mALb/Cr<300μg/mg) had 46 cases,the clinical albuminuria group(urinary mALb/Cr≥300μg/mg) included 30 cases,another 49 diabetic patients without kidney damage were seleted as control group.The urinary mALb/Cr,blood UA,fasting blood glucose(FBG),triacylglycerol(TG),total cholesterol(TC),high-density lipoprotein(HDL),low density lipoprotein(LDL),glycosylated hemoglobin(HbA1c) levels were determined.The BMI and the length of the course of the disease calculate.Results The patients' age,course of the disease,urinary mALb/Cr,blood UA,FBG,TC,TG,LDL,HbA1c and BMI level in the clinical albuminuria group and microalbuminuria group were significantly higher than those in the control group,the differences were statistically significant (F=6.18,12.48,141.43,12.48,8.49,4.98,6.18,3.89,3.17,3.89,all P<0.05).The high uric acid hematic disease rates of the clinical albuminuria group and microalbuminuria group were 26.09% and 26.09%,which were significantly higher than 10.20% of the control group,the differences were statistically significant(x2=4.074,24.833,all P<0.05).Urinary mALb/Cr was positively correlated with age,duration,BMI,UA,TG,TC,LDL,FBG,HbA1c(r=0.120,0.299,0.148,0.340,0.157,0.149,0.103,0.487,0.103).Multiple linear stepwise regression analysis suggested that duration,blood UA,FBG were independent risk factors of urinary mALb/Cr;TG,BMI,urinary mALb/Cr were independent risk factors for blood UA.Conclusion Urinary mALb/Cr and blood UA are the independent risk factors,high uric acid hematic disease may participate in the development process of DN,and diabetes duration,UA,BMI,TG,TC,LDL,FBG,HbA1C associated with increased urinary mALb/Cr excretory DN patients,the effective monitoring can improve the symptoms of DN and quality of life.

OBJECTIVETo 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).

METHODSGet 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.

RESULTSAfter 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.

CONCLUSIONSThe 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.

Adult ; Animals ; Dendritic Cells ; immunology ; Epitopes ; HLA-A2 Antigen ; immunology ; Humans ; K562 Cells ; Mice ; Peptides ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; Tumor Cells, Cultured ; alpha-Fetoproteins ; immunology

OBJECTIVETo establish a method for the prenatal diagnosis of 22q11 microdeletion syndrome.

METHODSBACs-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.

RESULTS22q11 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.

CONCLUSIONThe combination of prenatal BoBs and FISH can provide a method for the prenatal diagnosis of 22q11 microdeletion.

Adult ; Chromosome Deletion ; Chromosomes, Human, Pair 22 ; genetics ; DiGeorge Syndrome ; diagnosis ; embryology ; genetics ; Female ; Fetal Diseases ; diagnosis ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Pregnancy ; Prenatal Diagnosis