ABSTRACT
Doubling method is the technical barriers in maize haploid breeding. It was very important to establish the independent intellectual property rights for doubling method. In this experiment, the maize haploid inducer, TG15, was used for producing maternal haploids. Also, haploids were obtained from two kinds of maternal genotypes involved in the experiment, including high-oil type and common type. Significant differences were observed among offspring of various genotypes in the recovery of haploid fertilization. In 21 hybrid offspring haploids, the average powder rate was 8.28%, and the seed setting rate was 4.98%. The experimental results showed that when the hybrids were treated with 0.08% colchicine, the average powder rate and seed setting rate of offspring haploids were 35.53 and 20.30%, respectively, which were significantly higher than the hybrids with natural recovery ability. This study primarily established the doubling method of haploids called "bud seedling method" in China which was very practicably in maize doubled haploid breeding.
Subject(s)
Zea mays/genetics , Chromosomes, Plant , Colchicine/pharmacology , Genes, Plant , Genotype , Haploidy , Inflorescence/physiology , Plant Breeding/methods , Seeds/genetics , Zea mays/drug effectsABSTRACT
Doubled haploid (DH) technology, which is used for rapidly purifying genetic resources, is a key technology in modern maize breeding. The present study evaluated the tissue culture characteristics of maize haploid coleoptile sections, in order to provide a new way of haploid doubling. With 20 combinations of haploid coleoptile sections, obtained by hybridization within Reid, Tangsipingtou, and Term-tropical groups, as explants, we analyzed the induction and differentiation rate of callus, observed the number of root tip chromosomes in regenerated plants, and analyzed the pollen fertility. In addition, we used 47 SSR markers to analyze the genotypes of regenerated plants. The Reid and Tangsipingtou groups had significantly higher induction rates of haploid coleoptile callus compared to the Term-tropical group. Fifteen haploid plants were obtained which had 10 chromosomes in the root tips as assessed by I-KI staining. It was also noticed that the pollen of pollinated anthers were partially fertile. The haploid plants had genetic stability and showed no variation. The Reid and Tangsipingtou groups had good culture characteristics of haploid coleoptile sections, while the Term-tropical group had poor culture characteristics. Genotypes of haploid plants generated by tissue culture were evidenced to come from recombinant types of parents. Thus, this study established a tissue culture system of maize haploid coleoptile.
Subject(s)
Cotyledon/genetics , Haploidy , Zea mays/genetics , Chromosomes, Plant , Genomics , Genotype , Hybridization, Genetic , Microsatellite Repeats , Phenotype , Pollen , Regeneration , Tissue Culture TechniquesABSTRACT
ß-amyloid peptides (Aßs) can exert neurotoxic effects through induction of oxidative damage, whereas lipoic acid (LA), a powerful antioxidant, can alleviate oxidative damage. In this study, we explored the effect and mechanism of action of LA on beta-amyloid-intoxicated C6 glioma cells. Cells were randomly divided into three groups: control (vehicle), Aß, and LA + Aß. The LA + Aß group was treated with LA for 2 h, then both the Aß-only and the LA + Aß groups were incubated with 25 µM Aß for 24 h. Cell viability was measured by the MTT method. Mitochondrial reduced glutathione (GSH) and oxidized glutathione (GSSG) levels were detected by enzyme-linked immunosorbent assay (ELISA), and the GSH to GSSG ratio calculated. Real-time polymerase chain reaction and western blot analyses were used to detect MnSOD mRNA and protein, respectively. Aß significantly inhibited C6 cell proliferation compared with the control group (P < 0.05). LA markedly increased cell viability compared with the Aß group (P < 0.05). The increased GSSH and decreased GSH mitochondrial accumulation induced by Aß was profoundly reversed by treatment with LA (P < 0.05). Aß significantly reduced MnSOD expression compared to controls (P < 0.05), whereas LA pretreatment increased MnSOD expression compared with the Aß-only group (P < 0.05); MnSOD protein levels showed similar patterns. These results suggest that LA might protect Aß-intoxicated C6 glioma cells by alleviating oxidative damage, providing a new treatment strategy for neurodegenerative diseases.
Subject(s)
Amyloid beta-Peptides/metabolism , Antioxidants/pharmacology , Glioma/metabolism , Thioctic Acid/pharmacology , Amyloid beta-Peptides/toxicity , Animals , Cell Line, Tumor , Cell Proliferation/drug effects , Gene Expression , Glioma/genetics , Glutathione/metabolism , Glutathione Disulfide/metabolism , Mitochondria/drug effects , Mitochondria/metabolism , Oxidation-Reduction/drug effects , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rats , Superoxide Dismutase/genetics , Superoxide Dismutase/metabolismABSTRACT
This study aimed to discuss the effects of 3 different analgesia methods on serum IL-6 and IL-10 in patients after cesarean delivery. Thirty full-term women, who underwent cesarean delivery, were randomly assigned to 3 analgesia groups (10 cases each) as follows: intramuscular injection of 100 mg pethidine (NC group), patient controlled epidural analgesia (PCEA) of 5 mg morphine plus 150 mg ropivacaine (MR group), and patient controlled intravenous analgesia (PCIA) of 150 mg sufentanil plus 5 mg droperidol (SF group). An electronic analgesia pump was available in all 3 groups. At 4, 12, 24, and 48 h after surgery, visual analogue scale (VAS) pain scores were evaluated, IL-6 and IL-10 serum levels were measured, and adverse reactions were documented. The MR and SF groups responded well to analgesia. VAS scores at 12 and 24 h in these 2 groups were significantly lower than those in the NC group (P < 0.05). IL-6 and IL-10 levels were elevated to varying degrees postoperatively in all 3 groups. In the MR and SF groups, no significant difference occurred at each time point (P > 0.05), but compared with the NC group, significant differences were observed at 12 and 24 h (P < 0.05). Both PCIA and PCEA produced good analgesic effect, decreased postoperative level of serum IL-6, promoted release of anti-inflammatory factor IL-10, maintained balance in postoperative serum IL-6 level, and reduced the postoperative inflammatory response. Adverse reactions were significantly higher with epidural morphine than with intravenous sufentanil.
Subject(s)
Analgesia/methods , Cesarean Section , Interleukin-10/blood , Interleukin-6/blood , Analgesia/adverse effects , Female , Humans , PregnancyABSTRACT
This study aimed to investigate the value of magnetic resonance spectroscopy (MRS) imaging in assessing nasopharyngeal carcinoma radiotherapy during the early delayed reaction period. Eighty cases of nasopharyngeal cancer treated with radiotherapy within the same period underwent MRS imaging before or after radiotherapy. Of the 80 cases, 47 underwent MRS imaging on the 3rd, 4th, 6th, and 12th months after radiotherapy. The trends of the primary metabolite concentration at different time points were monitored and compared with the corresponding data after radiotherapy. Repeated measures analysis of variance was performed. At the end of radiotherapy, the N-acetyl aspartate (NAA)/creatine (Cr), choline (Cho)/Cr, and NAA/Cho ratios were reduced to the lowest levels after 3 months. However, increasing trends were observed from the 4th to the 12th month. On the 12th month, stable levels were reached with statistically significant differences (F = 316.02, 53.84, 286.68; P < 0.01). MRS reflected the radiation injury-repair process in the brain of a nasopharyngeal cancer patient during early delayed reaction. This non-invasive monitoring of changes in brain tissue metabolite concentrations provides valuable information for prognosis.
Subject(s)
Brain Injuries/diagnosis , Magnetic Resonance Imaging/methods , Magnetic Resonance Spectroscopy/methods , Nasopharyngeal Neoplasms/radiotherapy , Radiation Injuries/diagnosis , Adult , Aspartic Acid/analogs & derivatives , Aspartic Acid/metabolism , Brain Injuries/etiology , Brain Injuries/metabolism , Choline/metabolism , Creatine/metabolism , Female , Humans , Male , Middle Aged , Prognosis , Radiation Injuries/etiology , Radiation Injuries/metabolism , Radiotherapy/adverse effects , Radiotherapy/methods , Time FactorsABSTRACT
SRY-related high-mobility-group box 9 (Sox9) gene is a cartilage-specific transcription factor that plays essential roles in chondrocyte differentiation and cartilage formation. The aim of this study was to investigate the feasibility of genetic delivery of Sox9 to enhance chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells (hUC-MSCs). After they were isolated from human umbilical cord blood within 24 h after delivery of neonates, hUC-MSCs were untreated or transfected with a human Sox9-expressing plasmid or an empty vector. The cells were assessed for morphology and chondrogenic differentiation. The isolated cells with a fibroblast-like morphology in monolayer culture were positive for the MSC markers CD44, CD105, CD73, and CD90, but negative for the differentiation markers CD34, CD45, CD19, CD14, or major histocompatibility complex class II. Sox9 overexpression induced accumulation of sulfated proteoglycans, without altering the cellular morphology. Immunocytochemistry demonstrated that genetic delivery of Sox9 markedly enhanced the expression of aggrecan and type II collagen in hUC-MSCs compared with empty vector-transfected counterparts. Reverse transcription-polymerase chain reaction analysis further confirmed the elevation of aggrecan and type II collagen at the mRNA level in Sox9-transfected cells. Taken together, short-term Sox9 overexpression facilitates chondrogenesis of hUC-MSCs and may thus have potential implications in cartilage tissue engineering.
Subject(s)
Humans , Cell Differentiation/genetics , Chondrogenesis/genetics , Fetal Blood/cytology , Mesenchymal Stem Cells/cytology , SOX9 Transcription Factor/genetics , Aggrecans/biosynthesis , Blotting, Western , Cartilage/metabolism , Cell Proliferation/genetics , Chondrocytes/metabolism , Collagen Type II/biosynthesis , Flow Cytometry , Green Fluorescent Proteins , Gene Expression Regulation/physiology , Human Umbilical Vein Endothelial Cells/cytology , Immunohistochemistry , Immunophenotyping , Primary Cell Culture , Reverse Transcriptase Polymerase Chain Reaction , Tissue Engineering , TransfectionABSTRACT
SRY-related high-mobility-group box 9 (Sox9) gene is a cartilage-specific transcription factor that plays essential roles in chondrocyte differentiation and cartilage formation. The aim of this study was to investigate the feasibility of genetic delivery of Sox9 to enhance chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells (hUC-MSCs). After they were isolated from human umbilical cord blood within 24 h after delivery of neonates, hUC-MSCs were untreated or transfected with a human Sox9-expressing plasmid or an empty vector. The cells were assessed for morphology and chondrogenic differentiation. The isolated cells with a fibroblast-like morphology in monolayer culture were positive for the MSC markers CD44, CD105, CD73, and CD90, but negative for the differentiation markers CD34, CD45, CD19, CD14, or major histocompatibility complex class II. Sox9 overexpression induced accumulation of sulfated proteoglycans, without altering the cellular morphology. Immunocytochemistry demonstrated that genetic delivery of Sox9 markedly enhanced the expression of aggrecan and type II collagen in hUC-MSCs compared with empty vector-transfected counterparts. Reverse transcription-polymerase chain reaction analysis further confirmed the elevation of aggrecan and type II collagen at the mRNA level in Sox9-transfected cells. Taken together, short-term Sox9 overexpression facilitates chondrogenesis of hUC-MSCs and may thus have potential implications in cartilage tissue engineering.
Subject(s)
Cell Differentiation/genetics , Chondrogenesis/genetics , Fetal Blood/cytology , Mesenchymal Stem Cells/cytology , SOX9 Transcription Factor/genetics , Aggrecans/biosynthesis , Blotting, Western , Cartilage/metabolism , Cell Proliferation/genetics , Chondrocytes/metabolism , Collagen Type II/biosynthesis , Flow Cytometry , Gene Expression Regulation/physiology , Green Fluorescent Proteins , Human Umbilical Vein Endothelial Cells/cytology , Humans , Immunohistochemistry , Immunophenotyping , Primary Cell Culture , Reverse Transcriptase Polymerase Chain Reaction , Tissue Engineering , TransfectionABSTRACT
BALB/c mice with pulmonary tuberculosis (TB) develop a T helper cell type 1 that temporarily controls bacterial growth. Bacterial proliferation increases, accompanied by decreasing expression of interferon (IFN)-γ, tumour necrosis factor (TNF)-α and inducible nitric oxide synthase (iNOS). Activation of dendritic cells (DCs) is delayed. Intratracheal administration of only one dose of recombinant adenoviruses encoding granulocyte-macrophage colony-stimulating factor (AdGM-CSF) 1 day before Mycobacterium tuberculosis (Mtb) infection produced a significant decrease of pulmonary bacterial loads, higher activated DCs and increased expression of TNF-α, IFN-γ and iNOS. When AdGM-CSF was given in female mice B6D2F1 (C57BL/6J X DBA/2J) infected with a low Mtb dose to induce chronic infection similar to latent infection and corticosterone was used to induce reactivation, a very low bacilli burden in lungs was detected, and the same effect was observed in healthy mice co-housed with mice infected with mild and highly virulent bacteria in a model of transmissibility. Thus, GM-CSF is a significant cytokine in the immune protection against Mtb and gene therapy with AdGM-CSF increased protective immunity when administered in a single dose 1 day before Mtb infection in a model of progressive disease, and when used to prevent reactivation of latent infection or transmission.
Subject(s)
Adenoviridae/genetics , Granulocyte-Macrophage Colony-Stimulating Factor/genetics , Granulocyte-Macrophage Colony-Stimulating Factor/immunology , Tuberculosis, Pulmonary/therapy , Animals , Disease Models, Animal , Female , Genetic Therapy , Immunotherapy , Lung/metabolism , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Inbred DBA , Tuberculosis, Pulmonary/genetics , Tuberculosis, Pulmonary/immunology , Tuberculosis, Pulmonary/transmissionABSTRACT
OBJECTIVE: We compared early pulmonary (18)fluorodeoxyglucose ((18)FDG) uptake in infants who had very low birth weight with and without exposure to intrauterine inflammation by using positron emission tomography (PET). A secondary goal was to correlate (18)FDG uptake with later death or bronchopulmonary dysplasia. METHODS: Within 72 hours of birth, 22 singleton infants between 25 and 30 weeks of gestation had a thoracic PET scan after intravenous (18)FDG. Influx constants (K(i)) for (18)FDG were determined. Placental histology assessed exposure to intrauterine inflammation. RESULTS: Chorioamnionitis was found in 13 infants. Seven of these infants also had evidence of funisitis. No inflammation was detected in the remaining nine infants. Median (minimum, maximum) thoracic K(I) was 0.008 (0.006, 0.011) mL/min/mL in infants with funisitis, 0.006 (0.002, 0.008) in infants with chorioamnionitis only, and 0.006 (0.001, 0.015) in infants with no evidence of intrauterine inflammation (P=.16). No relation was found between K(i) and later death or bronchopulmonary dysplasia. Cord blood interleukin-6 was elevated in newborns with placental inflammation (P=.014). CONCLUSION: Early thoracic PET scanning for metabolically active inflammatory cells does not differ between infants with and without exposure to intrauterine inflammation. Evidence of early intrapulmonary sequestration of inflammatory cells in some infants without chorioamnionitis points to the complex etiology of postnatal inflammation.