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OBJECTIVES: This study assessed the protective effect of calcium dobesilate against contrast-induced nephropathy (CIN) after coronary angiography (CAG) or percutaneous coronary intervention (PCI) in patients with diabetes and chronic kidney disease (CKD). METHODS: A total of 130 patients with diabetes and CKD estimated glomerular filtration rate: 30-90 mL/min/1.73m2 were enrolled and included in the analysis. They were divided into experimental (n=65) and control groups (n=65). Patients in the experimental group were administered oral calcium dobesilate (500 mg) three times daily for 2 days before and 3 days after the procedure. The serum creatinine (SCr), cystatin C (Cys C), and neutrophil gelatinase-associated lipocalin (NGAL) levels were measured before and after the procedure. RESULTS: The mean SCr level at 24h after the procedure was found to be significantly lower in the experimental group than in the control group (79.1±19.6 µmol/L vs. 87.0±19.3 µmol/L, p=0.023). However, the Cys C and NGAL levels were not significantly different between the two groups at all measurement time points (all p>0.05). The incidence of CIN defined by the SCr level was significantly lower in the experimental group than in the control group (3 [4.6%] vs. 13 [20.0%], p=0.017). However, the incidence of CIN defined by the Cys C level was not statistically different between the two groups (7 [10.8%] vs. 7 [10.8%], p=1.000). CONCLUSIONS: This study revealed that calcium dobesilate has no preventive effect against CIN in patients with diabetes and CKD.
Assuntos
Dobesilato de Cálcio , Diabetes Mellitus , Nefropatias , Intervenção Coronária Percutânea , Insuficiência Renal Crônica , Biomarcadores , Meios de Contraste/efeitos adversos , Angiografia Coronária , Creatinina , Taxa de Filtração Glomerular , Humanos , Insuficiência Renal Crônica/complicaçõesRESUMO
PURPOSE: Esophageal cancer is a common malignant tumor in life, which seriously affects human health. In order to reduce the work intensity of doctors and improve detection accuracy, we proposed esophageal cancer detection using deep learning. The characteristics of deep learning: association and structure, activity and experience, essence and variation, migration and application, value and evaluation. METHOD: The improved Faster RCNN esophageal cancer detection in this paper introduces the online hard example mining (OHEM) mechanism into the system, and the experiment used 1520 gastrointestinal CT images from 421 patients. Then, we compare the overall performance of Inception-v2, Faster RCNN, and improved Faster RCNN through F-1 measure, mean average precision (mAP), and detection time. RESULTS: The experiment shows that the overall performance of the improved Faster RCNN is higher than the other two networks. The F-1 measure of our method reaches 95.71%, the mAP reaches 92.15%, and the detection time per CT is only 5.3s. CONCLUSION: Through comparative analysis on the esophageal cancer image data set, the experimental results show that the introduction of online hard example mining mechanism in the Faster RCNN algorithm can improve the detection accuracy to a certain extent.
Assuntos
Neoplasias Esofágicas , Redes Neurais de Computação , Algoritmos , Neoplasias Esofágicas/diagnóstico por imagem , Humanos , Tomografia Computadorizada por Raios XRESUMO
PURPOSE: In order to solve the problem of accurate and effective segmentation of the patient's lung computed tomography (CT) images, so as to improve the efficiency of treating lung cancer. METHOD: We propose a U-Net network (DC-U-Net) fused with dilated convolution, and compare the results of segmented lung CT with DC-U-Net, Otsu and region growth. We use Intersection over Union (IOU), Dice coefficient, Precision and Recall to evaluate the performance of the three algorithms. RESULTS: Compared with the common segmentation algorithm Otsu and region growing, the segmented image of DC-U-Net is closer to the Ground truth. The IOU of DC-U-Net is 0.9627, and the Dice coefficient is 0.9743, which is close to 1 and much higher than the other two algorithms. CONCLUSION: We propose that the model can directly segment the original image automatically, and the segmentation effect is good. This model speeds up the segmentation, simplifies the steps of medical image segmentation, and provides better segmentation for subsequent lung blood vessels, trachea and other tissues.
Assuntos
Processamento de Imagem Assistida por Computador , Neoplasias Pulmonares , Algoritmos , Humanos , Pulmão/diagnóstico por imagem , Neoplasias Pulmonares/diagnóstico por imagem , Tomografia Computadorizada por Raios XRESUMO
OBJECTIVES: This study assessed the protective effect of calcium dobesilate against contrast-induced nephropathy (CIN) after coronary angiography (CAG) or percutaneous coronary intervention (PCI) in patients with diabetes and chronic kidney disease (CKD). METHODS: A total of 130 patients with diabetes and CKD estimated glomerular filtration rate: 30-90 mL/min/1.73m2 were enrolled and included in the analysis. They were divided into experimental (n=65) and control groups (n=65). Patients in the experimental group were administered oral calcium dobesilate (500 mg) three times daily for 2 days before and 3 days after the procedure. The serum creatinine (SCr), cystatin C (Cys C), and neutrophil gelatinase-associated lipocalin (NGAL) levels were measured before and after the procedure. RESULTS: The mean SCr level at 24h after the procedure was found to be significantly lower in the experimental group than in the control group (79.1±19.6 μmol/L vs. 87.0±19.3 μmol/L, p=0.023). However, the Cys C and NGAL levels were not significantly different between the two groups at all measurement time points (all p>0.05). The incidence of CIN defined by the SCr level was significantly lower in the experimental group than in the control group (3 [4.6%] vs. 13 [20.0%], p=0.017). However, the incidence of CIN defined by the Cys C level was not statistically different between the two groups (7 [10.8%] vs. 7 [10.8%], p=1.000). CONCLUSIONS: This study revealed that calcium dobesilate has no preventive effect against CIN in patients with diabetes and CKD.
Assuntos
Humanos , Dobesilato de Cálcio , Diabetes Mellitus , Insuficiência Renal Crônica/complicações , Intervenção Coronária Percutânea , Nefropatias , Biomarcadores , Angiografia Coronária , Meios de Contraste/efeitos adversos , Creatinina , Taxa de Filtração GlomerularRESUMO
OBJECTIVE: To evaluate the efficacy and safety of low-dose rituximab in the treatment of hematologic abnormalities in patients with connective tissue disease. MATERIALS AND METHODS: A total of 13 patients with connective tissue disease who did not respond to prednisolone and multiple immunosuppressive agents, or their disease recurred after treatment, were given 100 mg of rituximab only combined with prednisolone once a week for 4 weeks. Then, the therapeutic effects and adverse reactions were respectively observed in the 13 patients. RESULTS: Rituximab showed good and rapid efficacy in the treatment of refractory thrombocytopenia and autoimmune hemolytic anemia caused by systemic lupus erythematosus, Sjögren's syndrome, and mixed connective tissue disease. Only 1 patient had urinary tract infection. During 24-month follow-up, disease recurred in 7 patients who still responded to azathioprine/Tripterygium wilfordii. CONCLUSION: Low-dose rituximab has good efficacy and safety in the treatment of hematologic abnormalities in patients with connective tissue disease.
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Doenças do Tecido Conjuntivo/tratamento farmacológico , Rituximab/uso terapêutico , Anemia Hemolítica Autoimune/tratamento farmacológico , Anemia Hemolítica Autoimune/etiologia , Anticorpos Monoclonais Murinos , Humanos , Lúpus Eritematoso Sistêmico/complicações , Síndrome de Sjogren/complicações , Trombocitopenia/tratamento farmacológico , Trombocitopenia/etiologia , Resultado do TratamentoRESUMO
Some patients have poor response to adult-onset Still disease (AOSD) traditional treatment, which easily recurs during the reduction of prednisone. We observed the efficacy and safety of tocilizumab combined with methotrexate (MTX) in the treatment of refractory AOSD, and to explore the possibility of reducing the dosage of tocilizumab after disease control.A total of 28 refractory AOSD cases who had an inadequate response to corticosteroids combined with at least 1 traditional immunosuppressive agent, and even large-dose prednisone could not relieve their conditions after recurrence, were selected in this study. They were treated with tocilizumab (intravenous 8âmg/kg) combined with MTX (oral 12.5âmg once a week). In detail, tocilizumab was firstly given every 4 weeks and after 6-month remission, it was then given every 8 weeks. Some items including body temperature, skin rash, joint swelling and pain, hepatosplenomegaly, blood routine, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), serum ferritin, and dosage of prednisone were observed before treatment as well as 2, 4, 8, 12, 24, 36, and 48 weeks after treatment. The adverse reactions occurring during the treatment were recorded.The body temperature was normal, the skin rash as well as joint swelling and pain disappeared, and laboratory indexes including CRP, ESR, white blood cell, neutrophilic granulocyte, platelet, hemoglobin, and ferritin were significantly improved after 8-week treatment (all Pâ<â.05). The clinical symptoms and laboratory indexes above mentioned were continuously improved 12, 24, 36, and 48 weeks after treatment. The mean dosage of prednisone was reduced from 71.4â±â20.7âmg/day to 55.0â±â11.1âmg/day after 2-week treatment, and to 3.3â±â2.1âmg/day after 48-week treatment (all Pâ<â.05). Prednisone was discontinued in 5 cases after 36-week treatment and in 7 cases after 48-week treatment. No serious adverse reactions occurred during the treatment.Tocilizumab can rapidly and markedly improve the clinical symptoms and laboratory indexes and contribute to reduction and discontinuation of prednisone in refractory AOSD. The patients' conditions are stable after reduction or discontinuation of prednisone and the tocilizumab possesses good safety.
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Anticorpos Monoclonais Humanizados/administração & dosagem , Antirreumáticos/administração & dosagem , Metotrexato/administração & dosagem , Doença de Still de Início Tardio/tratamento farmacológico , Administração Intravenosa , Administração Oral , Adulto , Anticorpos Monoclonais Humanizados/farmacologia , Antirreumáticos/farmacologia , Quimioterapia Combinada , Feminino , Glucocorticoides/administração & dosagem , Glucocorticoides/efeitos adversos , Humanos , Masculino , Metotrexato/farmacologia , Pessoa de Meia-Idade , Prednisona/administração & dosagem , Prednisona/efeitos adversos , Resultado do TratamentoRESUMO
Developing fluorescent probes for selective determination of the toxic and carcinogenic hydrazine are pretty significant. Herein, a rhodamine dye coupled to naphthalene was selected as a near-infrared fluorophore and acetyl group as a trigger unit for hydrazine sensing with a Stokes shifts of 62â¯nm. The probe showed about 77-fold NIR fluorescence enhancement in the presence of hydrazine. In addition, the detection limit was as low as 3.4â¯ppb, and the fluorescence intensity at 654â¯nm showed a satisfactory linearity with the concentration range of hydrazine from 0 to 120⯵M. More importantly, the practical utility of probe has been successfully proved through the fluorescence bioimaging of hydrazine in living cells with low cytotoxicity and quantitative N2H4 detection in environmental water samples.
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Corantes Fluorescentes/química , Hidrazinas/análise , Espectroscopia de Luz Próxima ao Infravermelho , Xantenos/química , Sobrevivência Celular , Células HeLa , Humanos , Concentração de Íons de Hidrogênio , Espectrometria de Fluorescência , Água/químicaRESUMO
Developing fluorescent probes for specific detection of extremely toxic thiophenols is pretty significant in the field of environment, chemistry and biology. We report herein a turn-on red fluorescent xanthene-based probe (RD-Probe) for detecting thiophenol with high selectivity over other analytes including aliphatic thiols. The probe could play the part of a "naked-eye"colorimetric indicator toward thiophenol. Moreover, the relative fluorescence intensity at 653â¯nm displayed good linearity with the concentration of thiophenol ranging from 0 to 6⯵M, and the limit of detection for thiophenol could be as low as 15â¯nM. Furthermore, the practicability of RD-Probe has been successfully proved through the quantitative thiophenol detection in real water samples and fluorescence bioimaging of thiophenol in HeLa cells.
Assuntos
Colorimetria , Corantes Fluorescentes/química , Imagem Óptica , Fenóis/análise , Compostos de Sulfidrila/análise , Poluentes Químicos da Água/análise , Xantenos/química , Corantes Fluorescentes/síntese química , Raios Infravermelhos , Estrutura Molecular , Espectrometria de Fluorescência , Espectrofotometria Ultravioleta , Xantenos/síntese químicaRESUMO
PURPOSE: This study investigated the impact of early enteral nutrition (EEN) on the clinical outcomes of gastric cancer patients after radical gastrectomy. METHODS: Four hundred gastric cancer patients undergoing radical gastrectomy of any extend with D2 nodal dissection were randomly divided into an experimental and a control group with 200 cases in each group. Patients in the control group received postoperative parenteral nutrition (PN), while patients in the experimental group received postoperative EEN. After treatment, the clinical outcomes, postoperative immune function, and nutritional status of the two groups were evaluated. RESULTS: The postoperative fever time, intestinal function recovery time, anal exhaust time, and the length of hospital stay for patients in the experimental group were significantly shorter than those of the control group. We did not find significant differences in anastomotic leak, postoperative ileus and regurgitation between the two groups. The activities of multiple immune cell types, including CD3âº, CD4âº, CD4âº/CD8âº, and natural killer (NK) cells, were significantly lower in both groups on postoperative day 1 when compared with the preoperative levels (p<0.05). The level of CD8⺠was not significantly different between the two groups (p>0.05). After treatment, levels of CD3âº, CD4âº, CD4âº/CD8âº, and NK cells in the experimental group patients were 35.6 ± 4.2, 42.2 ± 3.0, 1.7 ± 0.3, and 27.3 ± 5.3%, respectively, on postoperative day 7, which were similar to the preoperative levels. The immune cell levels from the control group patients remained significantly lower when compared with preoperative values; in addition, these values were also significantly lower when compared with the EEN patients (p<0.05) CONCLUSION: For gastric cancer patients undergoing radical gastrectomy, the clinical outcome, immune function and nutritional status after EEN were significantly improved. These data suggest the widespread use of EEN in clinical practice.
Assuntos
Nutrição Enteral/efeitos adversos , Neoplasias Gástricas/cirurgia , Adulto , Idoso , Feminino , Gastrectomia , Humanos , Masculino , Pessoa de Meia-Idade , Estado Nutricional , Neoplasias Gástricas/imunologiaRESUMO
OBJECTIVE: To investigate whether microsatellite instability (MSI) of gastric cancer and precancerous lesions were existed and its effect. METHODS: Laser microdissection was used. Gastric, intestinal metaplasia, dysplasia and normal mucosa were collected respectively. Five microsatellite loci were selected and MSI was detected by denaturing high-performance liquid chromatography. RESULTS: In the five microsatellite loci REF-positive phenotype, intestinal metaplasia MSI was 20.7%. Dysplasia MSI was 22.4%. Gastric MSI was 47.9%, and there was no MSI in normal gastric mucosa. CONCLUSION: MSI gradually increased from precancerous lesions to gastric cancer. The early detection of MSI may be a potential early warning indicator for early diagnosis of gastric cancer.
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OBJECTIVE: We aimed to investigate the association between gastric cancer and microsatellite instability (MSI) in the present study. METHOD: Phenol-chloroform method was employed for DNA extraction from the cancer tissues of 65gastric cancer patients and the dysplasia tissues and normal control tissues of 32 non-gastric cancer patients. The microsatellite loci Bat25, Bat26, D2S123, D5S346 and D17S250 were detected by using PCR-SSCP silver staining technique, and the MSI of the gastric cancer tissues and the precancerous tissues was analyzed. RESULTS: Of 65 gastric cancer cases, MSI was detected in 43 cases, with the detection rate of 66.2%. There were 13 cases showing MSI-H and 30 cases showing MSI-L, accounting for 30.2% and 69.8%, respectively. Among 32 cases of dysplasia tissues, MSI was detected in 10 cases, with the detection rate of 31.3%. Two cases of dysplasia tissues showed MSI-H and 8 cases showed MSI-L, accounting for 20.0% and 80.0%, respectively. CONCLUSION: Gastric cancer patients had a high detection rate of MSI. It is speculated that MSI is another molecular mechanism of carcinogenesis and may serve as a sensitive diagnostic indicator of gastric cancer.
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PURPOSE: Several studies indicated that the expression level of MLL3 gene in gastric cancer tissue was associated with prognosis, and previous studies also suggested that genetic polymorphisms of MLL3 were related to the risk for gastric cancer. The present study aimed to investigate the association of a missense mutation (S3660L) in the MLL3 gene with gastric cancer risk in a Chinese population. METHODS: In the present study, we identified a novel missense mutation in MLL3 gene (S3660L) by directly sequencing method in 48 gastric cancer patients. To further explore the relation between gastric cancer and this mutation, we selected 354 gastric cancer patients and 377 healthy control subjects and designed a case-control study. RESULTS: We found that the AG genotype (14.9 vs 6.40%, odds ratio/OR=2.58, 95% CI: 1.33-4.54, p<0.001) and A allele (7.5 vs 3.2%, OR=2.46, 95% CI: 1.55~5.34, p<0.001) were common in the gastric cancer patients than in the control subjects. CONCLUSION: We concluded that this novel missense (S3660L) mutation in MLL3 gene is likely to increase the gastric cancer risk.
Assuntos
Proteínas de Ligação a DNA/genética , Predisposição Genética para Doença , Mutação de Sentido Incorreto , Neoplasias Gástricas/genética , Adulto , Idoso , Estudos de Casos e Controles , Feminino , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , RiscoRESUMO
It is reported that the expression level of MLL3 in gastric cancer tissue highly correlates with tumor progression. However, whether MLL3 genetic variants are associated with the risk of gastric cancer remains unclear. In this study, we conducted a genotyping analysis for MLL3 in 314 cases of gastric cancer and 322 controls from the Chinese Han population. 4 SNPs (rs6943984, rs4725443, rs3800836, rs6464211) were selected for the present analysis. We found 2 SNPs (rs6943984, rs4725443) of MLL3 gene were significantly associated with the risk of gastric cancer : the rs6943984 with the minor allele A and rs4725443 with the minor allele C revealed strong associations with increased gastric cancer risk [P <0.001, OR=1.97, 95% CI=1.48~2.64 and P <0.001, OR = 2.23, 95% CI = 1.54~3.24]. Haplotype analysis of the four SNPs showed that haplotype A-T-A-C, G-T-G-C, and G-C-A-C increased the risk of gastric cancer (P <0.001, P=0.18, and P<0.001, respectively), while haplotype G-T-A-C significantly reduced the risk of gastric cancer (P <0.001). We concluded that MLL3 variants are significantly associated with gastric cancer risk. Our results for the first time provided new insight into susceptibility factors of MLL3 gene variants in carcinogenesis of gastric cancer of the Chinese Han population.
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Proteínas de Ligação a DNA/genética , Polimorfismo de Nucleotídeo Único/genética , Neoplasias Gástricas/etiologia , Estudos de Casos e Controles , China/epidemiologia , Feminino , Seguimentos , Predisposição Genética para Doença , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Fatores de Risco , Neoplasias Gástricas/epidemiologiaRESUMO
OBJECTIVE: To determine the blood-sucking habit of anopheline by mosquitoes bloodmeal identification in malaria endemic area Chayu County, Tibet Autonomous Region. METHODS: Three villages with different bio-environments were selected as the investigation spots. Light traps were set up outdoor and in livestock sheds from sunset (20:00) to sunrise (8:00) in 3-4 consecutive nights to collect mosquitoes. The trapped anophelines were counted and identified according to morphological criteria and multiple PCR method. A PCR-based methodology according to the mtDNA-cytb variations was used in different mammal hosts to identify bloodmeal sources in engorged mosquitoes. The human blood index (HBI) was assessed to determine the range of hosts. RESULTS: Among 1 442 anopheline mosquitoes collected by 108 light traps on 13 nights. 1436 (99.6%) belonged to Anopheles maculatus complex, with 85.5% An. pseudo-willmori and 14.5% An. willmori. Positive bloodmeal identification was found from 168 (83%) of 202 field-collected engorged mosquitoes. The crude HBI of An. pseudowillmori and An. willmori were 035 and 0.29, respectively. CONCLUSION: An. pseudowillmori and An willmori are both zoophilic and anthropophilic, and An. pseudowillmori shows a higher HBI.
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Anopheles/fisiologia , Anopheles/parasitologia , Malária/transmissão , Reação em Cadeia da Polimerase Multiplex , Animais , Comportamento Alimentar , Humanos , Malária/epidemiologia , Tibet/epidemiologiaRESUMO
BACKGROUND: Liver revascularization is frequently required during the enlarged radical operation for hilar cholangiocarcinoma involving the hepatic artery. Researchers have carried out a number of experiments applying partial portal vein arterialization (PVA) in clinical practice. In this study we aimed to establish a theoretical basis for clinical application of partial PVA and to investigate the effects of partial PVA on rat hilar bile duct and hepatic functions. METHODS: Thirty rats were randomly and equally assigned into 3 groups: control (group A), hepatic artery ligation+bile duct recanalization (group B), and partial PVA+bile duct recanalization (group C). Proliferation and apoptosis of rat hilar bile duct epithelial cells, arteriolar counts of the peribiliary plexus (PBP) of the bile duct wall, changes in serum biochemistry, and pathologic changes in the bile duct were assessed 1 month after operation. RESULTS: The proliferation of hilar bile duct epithelial cells in group B was greater than in groups A and C (P<0.01). No apoptotic hilar bile duct epithelial cells were detected in any of the groups. The PBP arteriolar counts of the hilar bile duct wall were similar in groups A and C (P>0.05), but the count was lower in group B than in group A (P<0.01). No statistically significant differences in alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and albumin were found in the 3 groups. The gamma-glutamyltransferase value was higher in group B than in groups A and C (P<0.01). The hepatic tissues of groups A and C showed no significant abnormality. Chronic inflammatory changes in the hilar bile duct walls were observed only in group B. CONCLUSION: Partial PVA can restore the arterial blood supply of the hilar bile duct and significantly extenuate the injury to hilar bile duct epithelial cells resulting from hepatic artery ligation.
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Ductos Biliares Intra-Hepáticos/irrigação sanguínea , Artéria Hepática/cirurgia , Fígado/irrigação sanguínea , Veia Porta/cirurgia , Animais , Apoptose , Ductos Biliares Intra-Hepáticos/metabolismo , Ductos Biliares Intra-Hepáticos/patologia , Biomarcadores/sangue , Proliferação de Células , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Ligadura , Fígado/metabolismo , Fígado/patologia , Testes de Função Hepática , Masculino , Modelos Animais , Ratos , Ratos Sprague-Dawley , Fatores de TempoRESUMO
OBJECTIVE: To investigate the current status of disabled children and prevalence of disabilities in children aged 0-6 years and their risk factors, and to provide scientific evidence for making relevant policies for disabled children. METHODS: In a community-based cross-sectional study, multi-phase, stratified, unequal proportional and cluster sampling was adopted to survey 60 124 children aged 0-6 years. All the investigated children were screened for disabilities, and those with positive screening tests were further diagnosed by various specialties. RESULTS: A total of 819 children were diagnosed as disabled with an overall prevalence of 1.362%, 0.155% for hearing disability, 0.160% for visual disability, 0.931% for intelligent disability, 0.424% for limb disability, and 0.101% for mental disability. Prevalence of disability in children was higher in rural areas, and in families with two or more children, low educational level or in divorced families. CONCLUSION: The prevalence of disability can be reduced by economic development, improvement of health care and quality of population, as well as harmonious familial relationship, early prevention of disability, and preschool education for disabled children.
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Cegueira/epidemiologia , Perda Auditiva/epidemiologia , Deficiência Intelectual/epidemiologia , Deformidades Congênitas dos Membros/epidemiologia , Criança , Pré-Escolar , China/epidemiologia , Pessoas com Deficiência , Humanos , Lactente , Recém-NascidoRESUMO
Agrobacterium-mediated transformation in chrysanthemum was studied to prevent the insect pest of aphid (Mizus persicae). The gna gene was successfully transferred into chrysanthemum by leaf dish, and 93 transgenic clones were obtained. The highest transformation frequency 11.21% was achieved on the optimization facts, which were medium YEB with pH5.6, bacterial concentration OD600 = 0.4, precultivation for one day, cocultivation for four days, the cocultivation media supplemented with GA3 0.5 mg/L and leaf explants growed for 45 days. The results from PCR and FQ-PCR analysis confirmed that gna gene was integrated into the genome of chrysanthemum plants. The insect bioassay with aphid showed that the aphid resistance of different transgenic plants was difference, and the rate of aphid population inhibition of them were from 10% to 84% with an average rate of 39.4%. The leaf-extracts from different transgenic plants showed varying actinties in red-blood cell bioassay.