ABSTRACT
Haploids and doubled haploids are critical components of plant breeding. This review is focused on studies on haploids and double haploids inducted in cucurbits through in vitro pollination with irradiated pollen, unfertilized ovule/ovary culture, and anther/microspore culture during the last 30 years, as well as comprehensive analysis of the main factors of each process and comparison between chromosome doubling and ploidy identification methods, with special focus on the application of double haploids in plant breeding and genetics. This review identifies existing problems affecting the efficiency of androgenesis, gynogenesis, and parthenogenesis in cucurbit species. Donor plant genotypes and surrounding environments, developmental stages of explants, culture media, stress factors, and chromosome doubling and ploidy identification are compared at length and discussed as methodologies and protocols for androgenesis, gynogenesis, and parthenogenesis in haploid and double haploid production technologies.
Subject(s)
Cucurbitaceae/physiology , Gametogenesis, Plant , Haploidy , Parthenogenesis , Chromosomes, Plant/genetics , Pollination/physiologyABSTRACT
AIM: To investigate the effects of our tumor vaccines on reversing immune tolerance and generating therapeutic response. METHODS: Vaccines were synthesized by solid phase using an Fmoc strategy, where a small molecule toll-like receptor-7 agonist (T7) was conjugated to a monoclonal gastric cancer 7 antigen mono-epitope (T7-MG1) or tri-epitope (T7-MG3). Cytokines were measured in both mouse bone marrow dendritic cells and mouse spleen lymphocytes after exposed to the vaccines. BALB/c mice were intraperitoneally immunized with the vaccines every 2 wk for a total of three times, and then subcutaneously challenged with Ehrlich ascites carcinoma (EAC) cells. Three weeks later, the mice were killed, and the tumors were surgically removed and weighed. Serum samples were collected from the mice, and antibody titers were determined by ELISA using an alkaline phosphate-conjugated detection antibody for total IgG. Antibody-dependent cell-mediated cytotoxicity was detected by the lactate dehydrogenase method using natural killer cells as effectors and antibody-labeled EAC cells as targets. Cytotoxic T lymphocyte activities were also detected by the lactate dehydrogenase method using lymphocytes as effectors and EAC cells as targets. RESULTS: Vaccines were successfully synthesized and validated by analytical high performance liquid chromatography and electrospray mass spectrometry, including T7, T7-MG1, and T7-MG3. Rapid inductions of tumor necrosis factor-α and interleukin-12 in bone marrow dendritic cells and interferon γ and interleukin-12 in lymphocytes occurred in vitro after T7, T7-MG1, and T7-MG3 treatment. Immunization with T7-MG3 reduced the EAC tumor burden in BALB/c mice to 62.64% ± 5.55% compared with PBS control (P < 0.01). Six or nine weeks after the first immunization, the monoclonal gastric cancer 7 antigen antibody increased significantly in the T7-MG3 group compared with the PBS control (P < 0.01). As for antibody-dependent cell-mediated cytotoxicity, antisera obtained by immunization with T7-MG3 were able to markedly enhance cell lysis compared to PBS control (31.58% ± 2.94% vs 18.02% ± 2.26%; P < 0.01). As for cytotoxic T lymphocytes, T7-MG3 exhibited obviously greater cytotoxicity compared with PBS control (40.92% ± 4.38% vs 16.29% ± 1.90%; P < 0.01). CONCLUSION: A successful method is confirmed for the design of gastric cancer vaccines by chemical conjugation of T7 and multi-repeat-epitope of monoclonal gastric cancer 7 antigen.
Subject(s)
Antigens, Neoplasm/immunology , Cancer Vaccines/pharmacology , Carcinoma, Ehrlich Tumor/drug therapy , Immunoconjugates/pharmacology , Membrane Glycoproteins/agonists , Toll-Like Receptor 7/agonists , Tumor Escape/drug effects , Animals , Antibody-Dependent Cell Cytotoxicity , Cancer Vaccines/administration & dosage , Cancer Vaccines/chemical synthesis , Cancer Vaccines/immunology , Carcinoma, Ehrlich Tumor/immunology , Carcinoma, Ehrlich Tumor/pathology , Cells, Cultured , Cytokines/metabolism , Epitopes , Female , Immunization Schedule , Immunoconjugates/administration & dosage , Injections, Intraperitoneal , Lymphocytes, Tumor-Infiltrating/drug effects , Lymphocytes, Tumor-Infiltrating/immunology , Lymphocytes, Tumor-Infiltrating/metabolism , Membrane Glycoproteins/immunology , Membrane Glycoproteins/metabolism , Mice, Inbred BALB C , Signal Transduction/drug effects , Superantigens , T-Lymphocytes, Cytotoxic/drug effects , T-Lymphocytes, Cytotoxic/immunology , T-Lymphocytes, Cytotoxic/metabolism , Time Factors , Toll-Like Receptor 7/immunology , Toll-Like Receptor 7/metabolism , Tumor BurdenABSTRACT
OBJECTIVE: To investigate the changes of physiological activities and functions in vitro of apheresis platelets during storage. METHOD: 17 units of apheresis platelets were randomly chosen and stored at 20 °C to 24 °C with agitation. Platelet counting (Plt), mean platelet volume (MPV), blood gases, pH value, glucose (Glu) concentration, lactate (LA) concentration, LDH concentration, thromboelastogram (TEG), hypotonic shock response (HSR), CD62p expression rate and anew expression rate were measured on days 0, 1, 3, 5 after platelet storage. Changes of physiological activities and functions in vitro were systematically evaluated by above-mentioned indexes. RESULTS: During storage, Plt, MPV and HSR were not significantly changed; but pH value, blood gases, Glu, LA, LDH, HSR, expression rate of CD62p and anew expression rate were significant differenty. Among thromboelastogram indexes, R value increased obviously with prolongation of storage time; K value and αAngle were not significantly changed; MA was not significantly changed on day 1 and 3, but was slightly increase on day 5. CONCLUSION: The physiological activities and functions in vitro of apheresis platelets are kept well during storage. For clinical transfusion of apheresis platelet during storage, clinical effect of transfusione is not influenced.