Bacterial canker caused by Pseudomonas syringae pv. Actinidiae is one of the most important diseases of kiwifruit (Actinidia chinensis) and leads to considerable yield losses. In order to obtain transgenic plants with resistance for 'Red Sun' kiwifruit to canker disease, a non-specific lipid transfer protein-like antimicrobial protein gene (LJAMP2) from motherwort (Leonurus japonicus) was introduced into 'Red Sun' kiwifruit through Agrobacterium-mediated transformation. After two days of co-cultivation with A. tumefaciens strain LBA4404 harboring 35S:LJAMP2, the transformed explants were transferred to the selection medium containing 25 mg/L kanamycin+3.0 mg/L BA+1.0 mg/L NAA. The regeneration efficiency of kanamycin-resistant shoots reached to 85%. All (100%) of kanamycin-resistant shoots rooted on half-strength MS medium supplemented with 0.8 mg/L IBA and a total of 40 regenerated plantlets were obtained. PCR and histochemical GUS activity analysis show that 23 of 40 lines (57.50%) were positive, suggesting that the LJAMP2 gene was integrated into the genome of 'Red Sun' kiwifruit. Taken together, we established an efficient genetic transformation method for 'Red Sun' kiwifruit using A. tumefaciens and the transformation frequency reached 5.11%. This protocol will be useful for the genetic breeding of 'Red Sun' kiwifruit for improvement of disease resistance.
Actinidia ; genetics ; Agrobacterium ; Antigens, Plant ; genetics ; Carrier Proteins ; genetics ; Leonurus ; Plant Proteins ; genetics ; Plants, Genetically Modified ; genetics ; Transformation, Genetic

OBJECTIVETo synthesize artificial antigen of the chlorogenic acid (CGA), a constituent in Traditional Chinese Medicine, then optimize the reaction conditions, so that to supply the basis in material and technique for study on Chinese medicine injection of adverse reactions in-depth.

METHODThe complete antigen of chlorogenic acid (CGA-BSA) was synthesized by CGA coupled to carrier protein bovine serum albumi (BSA) using carbodiimide method. We used the orthogonal design test to optimize reaction conditions and used the MALDI-TOF-MS and UV motheds to determine the coupling rate of the CGA-BSA.

RESULTThe coupling rate were 20 and 18.8, counting by MALDI-TOF-MS and UV, respectively; while the best reaction conditions were NHS: CGA, EDC: CGA, CGA: BSA were 4: 1, 1: 1, 200: 1, respectively, with the pH 9.

CONCLUSIONThe complete antigen is preparated successfully. Which supplied the basis to advanced study of correlation.

Antigens, Plant ; chemistry ; Chlorogenic Acid ; chemical synthesis ; chemistry ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Spectrophotometry, Ultraviolet

The objective of this study was to explore the effect of astragalus polysaccharide (APS) on sensitivity of leukemic cell line HL-60 to NK cell cytotoxicity and its mechanism. The cytotoxicities of NK cells against HL-60 cells were analyzed by LDH releasing assay at different effect-to-target cell ratios (E:T) before and after treated with APS. The gene expression of MHC class I chain-related (MICA) in HL-60 cells before and after APS treatment was assayed with RT-PCR. Protein expression of MICA in HL-60 cells was assayed by flow cytometry before and after treated by APS. The results showed that after treated with APS 15 mg/ml for 48 h, the cytotoxicities of NK cells against HL-60 cells enhanced at different effect-to-target (P < 0.05), and the gene and protein expressions in MICA of HL-60 cells were up-regulated (P < 0.05). It is concluded that the APS can obviously up-regulate the expression of MICA in HL-60 cells, thus enhance sensitivity of HL-60 cells to cytotoxicity of NK cells.
Astragalus Plant ; Cytotoxicity, Immunologic ; drug effects ; HL-60 Cells ; Histocompatibility Antigens Class I ; metabolism ; Humans ; Killer Cells, Natural ; Polysaccharides ; pharmacology

Peanut is one of the most popular foods in the world due to its high nutrition; however, it contains multiple seed storage proteins which are identified as allergens and hence are the most common cause of life-threatening, IgE-mediated anaphylaxis among the hypersensitive individuals. Three peanut proteins, Arachis hypogaea allergy 1, 2, 3 (Ara h 1, Ara h 2 and Ara h 3), which have the common biochemical characteristics like resistance to proteases and heat, are considered as the major allergens because they are recognized by serum IgE from a peanut-allergic patient population. The linear IgE-binding epitopes in the allergens lay the foundation of the anaphylaxis in the peanut-allergic individuals. Peanut allergy is often a life-long problem, so many investigators are focusing on decreasing clinical reactivity. In this review, the latest advances in the researches on biochemical characteristics, structure and function of the three major allergens were described and particular attention was given to the immunity properties of the three allergens. The future research directions were also discussed.
2S Albumins, Plant ; chemistry ; genetics ; immunology ; Animals ; Antigens, Plant ; chemistry ; genetics ; immunology ; Arachis ; chemistry ; DNA ; genetics ; Glycoproteins ; chemistry ; genetics ; immunology ; Humans ; Immunoglobulin E ; genetics ; Plant Proteins ; chemistry ; genetics ; immunology

Pollinosis is one of the allergic diseases which caused by airborne pollens from allergic plants, such as allergic rhinitis, asthma, and allergic conjunctivitis. The distribution of pollens in the air is fully related to weather condition, especially temperature and humidity, etc. This study is focused on the evaluation of the relation between pollen count and the outbreak of allergic diseases (symptom index) from allergic patients in Korea and this data would be necessary to evaluate Korean own allergy index for pollen forecasting system. Airborne particles carrying allergens were collected daily from nationwide 8 stations (Seoul, Guri, Daejeon, Daegu, Kwangju, Busan, Kangneung, and Jeju) by using 7 days-Burkard sampler (Burkard Manufacturing Co Ltd, Hertfordshire, UK) for 10 year in South Korea (March 1, 2008-October 30, 2008). They were counted and recorded along with the weather factors. Symptom index was evaluated and recorded by calling to allergic patients daily. Airborne pollen has two peak seasons that is May and September in Korea. The peak season for mold was June. There was the tree pollen seasons from the middle of February to late July, then followed during the grasses and weed season from the middle of July to the end of October. Ragweed was one of the most common pollen of autumn in Korea. In skin prick test, Ragweed was the highest sensitization rate (5.9%), followed by Mugwort (5.8%), alder (4.9%), birch (4.8%), oak grasses among the pollen extracts in Korean school aged children. Ragweed pollen counts were gradually increased since 2000. In same period, sensitization rates for airborne pollen, especially weeds in Korean children were increased annually (1999: 4.4%, 2000: 7.1%, 2001: 7.4%, 2002: 8.5%, 2003: 7.7%). There are significantly related between symptom index from allergic patients and allergic pollen concentrations. Sensitization rates for pollens are increased in Korean children with age. There are significantly related between symptom index from allergic patients and allergic pollen concentrations. Risk Grade of pollen for allergy pollen forecast among each allergic plant was established and still remain to confirm it.
Aged ; Allergens ; Alnus ; Ambrosia ; Antigens, Plant ; Artemisia ; Asthma ; Betula ; Child ; Conjunctivitis, Allergic ; Forecasting ; Fungi ; Humans ; Humidity ; Hypersensitivity ; Korea ; Lifting ; Plant Extracts ; Plants ; Poaceae ; Pollen ; Republic of Korea ; Rhinitis ; Rhinitis, Allergic, Perennial ; Rhinitis, Allergic, Seasonal ; Seasons ; Skin ; Weather

Ex vivo maintainance of human stem cells is crucial for many clinical applications. Current culture conditions provide some level support but cytokines induce most quiescent stem cells to proliferate and differentiate. Better control of primitive cells is needed to extend the time and range of manipulation of such cells. A recently identified plant lectin Flt3 receptor-interacting lectin (FRIL) present may a special ability to preserve primitive CB progenitors for extended periods in culture without exogenous cytokines. But the mechanisms of FRIL preserving quiescent primitive cells are still unknown. Recently a novel protein HTm4 and its alternatively spliced variant HTm4S, which serve as hematopoietic cell cycle regulators, have been identified. In this report we studied the effect of FRIL on the in vitro maintenance of quiescent human cord blood stem cells and the expression of the novel hematopoietic cell cycle regulator HTm4 and HTm4S in progenitor cells cultured in FRIL. We analyzed the proliferation and the HPP-CFC proportion of CD34(+) cells treated with FRIL. The human HTm4 and HTm4S mRNA expression was detected by semi-quantitative RT-PCR, and the cell cycle status of CB CD34(+) cells was analyzed by FACS. The results showed that incubation of CD34(+) cells in FRIL resulted in a low proliferation of progenitor cells and fewer cycling cells, but FRIL selectively maintained a higher number of primitive cells with proliferative potential in suspension culture. CB CD34(+) cells cultured in FRIL showed significant diversity in the expression of HTm4 and HTm4S during 0~14 d. On d 0, HTm4 was detected at high level, downregulated on d 1, but upregulated during d 3 to d 14, and reaching the highest level on d 7. But the expression levels of HTm4S changed little in the cells cultured in FRIL except the obviously increased expression on d 7. Exogenous expression showed that HTm4 was localized around the karyon while HTm4S scatted in the cytoplasm, respectively, which may be responsible for their difference in function. Thus, FRIL can preserve quiescent primitive CD34(+), and FRIL's ability to preserve quiescent primitive cells in a reversible manner may significantly expand the time and range of ex vivo manipulations of human stem cells for clinical applications. In other words, HTm4 and HTm4S may play a crucial role in the cell cycle modulation of CD34(+) progenitor cells maintained with FRIL in vitro.
Antigens, CD20 ; biosynthesis ; genetics ; Antigens, CD34 ; metabolism ; Cell Cycle ; Cell Cycle Proteins ; biosynthesis ; genetics ; Cell Separation ; Cells, Cultured ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans ; Mannose-Binding Lectins ; pharmacology ; Membrane Proteins ; biosynthesis ; genetics ; Plant Lectins ; pharmacology

OBJECTIVETo explore the activities of Composite Artemisia Capillaris Tablet (CACT) against hepatitis B virus replication in vitro.

METHODSBy means of radioimmunoassay (RIA), Dot blot and Southern blot, the surface and e antigen production of 2.2.15 cells, HBV DNA in 2.2.15 cell culture medium and that in 2.2.15 cells were examined respectively.

RESULTSHBsAg, HBeAg values of 2.2.15 cells treated by CACT were lower than those of the control, the HBV DNA quantities in culture medium and in 2.2.15 cells decreased as compared with those cells with no treatment by CACT given to them.

CONCLUSIONCACT could inhibit HBV DNA replication, showing its potential antiviral activity in hepatitis B treatment.

Cell Line, Tumor ; DNA, Viral ; antagonists & inhibitors ; Hepatitis B Surface Antigens ; metabolism ; Hepatitis B e Antigens ; metabolism ; Hepatitis B virus ; genetics ; immunology ; physiology ; Humans ; Medicine, Chinese Traditional ; Plant Preparations ; administration & dosage ; pharmacology ; toxicity ; Radioimmunoassay ; Tablets ; Virus Replication ; drug effects

By now, the digestive stability experiments provided by most authoritative organizations are in vitro tests. Evaluating the protein digestive stability with in vivo models should be more objective. The present study aimed to verify the in vivo digestibility of soybean β-conglycinin β-subunit in Wuzhishan (WZS) minipigs. Three minipigs were surgically fitted with O-stomach and T-ileum cannulae and fed with soybean meals. According to SDS-PAGE, the 50 kD fraction of soybean β-conglycinin β-subunit persisted in the gastric fluid until 6 h after feeding, which was detected at 3 h and clearly visible at 4-6 h in the intestinal fluid. Western blot with anti-β-conglycinin β-subunit McAb confirmed it.
Animals ; Antigens, Plant ; chemistry ; metabolism ; Digestion ; physiology ; Globulins ; chemistry ; metabolism ; Male ; Protein Subunits ; chemistry ; metabolism ; Seed Storage Proteins ; chemistry ; metabolism ; Soybean Proteins ; chemistry ; metabolism ; Swine ; Swine, Miniature ; physiology

OBJECTIVETo investigate the anti-atherosclerosis mechanism of Astragalus mongholicus polysaccharides (APS) by examining its effect on gene expression profiles of the dendritic cells (DCs) from healthy donors.

METHODSPeripheral blood DCs from healthy donors were incubated with 200 mg/L APS overnight, and changes in the gene expression profiles were investigated using microarray technique and RT-PCR.

RESULTSCompared with the control cells, APS-treated DCs showed significantly up-regulated expressions of CD36 (0.97 ± 0.23 vs 5.45 ± 1.14) and IL-27 (1.08 ± 0.22 vs 2.97 ± 0.61) and down-regulated expression of expression of IFI16 (0.98 ± 0.18 vs 0.46 ± 0.11).

CONCLUSIONSAPS can promote the maturation and differentiation of DCs by up-regulating CD36 and IL-27 and down-regulating IFI16, and thus positively affects the occurrence and progression of the atherosclerosis.

Astragalus Plant ; chemistry ; CD36 Antigens ; metabolism ; Cell Differentiation ; Dendritic Cells ; drug effects ; Humans ; Interleukins ; metabolism ; Nuclear Proteins ; metabolism ; Phosphoproteins ; metabolism ; Polysaccharides ; pharmacology ; Transcriptome

OBJECTIVETo establish WZS miniature swine model of beta-conglycinin (7 S) allergy for evaluating the potential allergenicity of genetically modified food.

METHODSTwelve 45-day-old WZS miniature swines from three litters were randomly divided into three groups (control group; 4% 7 S group and 8% 7 S group, n = 4), which were respectively gastric sensitized (day 0 - 10) and oral challenged (day 6 - 18, 31) to induce anaphylactic reactions. Clinical symptoms, skin prick reactions were recorded. At day 10, 19 and 32, serum IgG, IgE, histamine and cytokines levels were measured by ELISA.

RESULTSDiarrhea at different degrees were observed in 4% and 8% 7 S groups. The skin erythema reactions in grade "-", "+/-", "+", "++" of control group respectively were 2/4, 2/4, 0/4, 0/4, of 4% 7 S group were 0/4, 0/4, 2/4, 2/4 and of 8% 7 S group were 0/4, 0/4, 1/4, 3/4. The serum IgE and histamine levels of day 11, 19 and 32 were all significantly and positively correlated (Pearson coefficients = 1, P = 0.000). The serum IgG, IgE and histamine levels all reached the peak after 7 S groups were oral challenged at day 19.Compared with the control group, serum IgG (lg IgG: 2.95 +/- 0.31 vs 2.29 +/- 0.25, t = 3.19, P = 0.011), IgE (lg IgE: 2.45 +/- 0.30 vs 1.77 +/- 0.23, t = 3.31, P = 0.009) and histamine levels(lg histamine:2.13 +/- 0.30 vs 1.45 +/- 0.23, t = 3.34, P = 0.009) of 4% 7 S group at day 19 were all significantly higher, while the serum IFN-gamma content [(35.78 +/- 6.42) pg/ml vs (51.10 +/- 9.67) pg/ml, t = -2.33, P = 0.045] of 4%7 S group was significantly lower.

CONCLUSIONThe WZS miniature swine model orally induced by soybean beta-conglycinin is type I hypersensitivity mediated by IgE, which can be used to predict the potential allergenicity of genetically modified food.

Animals ; Antigens, Plant ; adverse effects ; Disease Models, Animal ; Food Hypersensitivity ; etiology ; Globulins ; adverse effects ; Seed Storage Proteins ; adverse effects ; Soybean Proteins ; adverse effects ; Swine ; Swine, Miniature