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1.
Br J Radiol ; 96(1147): 20220492, 2023 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-37066834

RESUMO

OBJECTIVE: To evaluate correlation between contrast-enhanced ultrasonography Liver Imaging Reporting and Data System (CEUS LI-RADS; v. 2017) categories (LR 3-5 vs LR-M) and outcomes in patients with early-stage hepatocellular carcinoma (HCC) after initial therapy. METHODS: In this retrospective study, 272 patients with high risks for HCC and solitary clinically or pathologically confirmed HCC were identified between January 2010 and December 2015. Patients were initially treated by resection and radiofrequency ablation (RFA) according to the Barcelona Clinic Liver Cancer staging system and were followed up until December 31, 2018. Recurrence-free survival (RFS) and overall survival (OS) were compared between nodules assigned as LR 3-5 or LR M according to CEUS LI-RADS v. 2017 by using the Kaplan-Meier curve, log-rank test, and Cox proportional hazard model. RESULTS: Early washout is the key determinating whether a nodule is classed as LR-M. Treatment procedures and LI-RADS category showed an independent correlation with OS and RFS (p < 0.05). LR 3-5 category were more correlated with better OS (88.6 months and 74.2 months, respectively; p = 0.017) compared with LR-M. Surgical resection demonstrated longer OS and RFS than RFA in LR-M patients and longer OS in LR 3-5 patients (p < 0.05). Besides, there was no significantly difference in OS and RFS between two categories in resection (p > 0.05), while for patients treated with RFA, LR 3-5 patients showed significant longer OS and RFS than LR-M patients (p < 0.05). CONCLUSION: Patients with HCC assigned as LR-M showed worse RFS and OS and surgical resection tended to be a more effective treatment for these patients. ADVANCES IN KNOWLEDGE: Putting forward a theory that CEUS LI-RADS categories could independently predict the outcome for patients with solitary HCC at early-stage after initial treatment.


Assuntos
Carcinoma Hepatocelular , Neoplasias Hepáticas , Humanos , Carcinoma Hepatocelular/diagnóstico por imagem , Carcinoma Hepatocelular/cirurgia , Neoplasias Hepáticas/diagnóstico por imagem , Neoplasias Hepáticas/cirurgia , Estudos Retrospectivos , Meios de Contraste , Imageamento por Ressonância Magnética/métodos , Ultrassonografia/métodos , Sensibilidade e Especificidade
2.
Bioconjug Chem ; 34(6): 1045-1053, 2023 06 21.
Artigo em Inglês | MEDLINE | ID: mdl-37097628

RESUMO

Here, lipase encapsulation is constructed by locking enzyme molecules in nanomolecular cages on the surface of SH-PEI@PVAC magnetic microspheres. To improve the encapsulation efficiency in enzyme loading, the thiol group is efficiently modified on the grafted polyethyleneimine (PEI) using 3-mercaptopropionic acid. N2 adsorption-desorption isotherms reveal the existence of mesoporous molecular cages on the microsphere surface. The robust immobilizing strength of carriers to lipase demonstrates the successful encapsulation of enzymes in nanomolecular cages. The encapsulated lipase shows high enzyme loading (52.9 mg/g) and high activity (51.4 U/mg). Different sizes of molecular cages are established, and the cage size showed important effects on lipase encapsulation. It shows that enzyme loading is low at a small size of molecular cages, which is attributed to that the nanomolecular cage is too small to house lipase. The investigation in lipase conformation suggests that the encapsulated lipase retains its active conformation. Compared with the adsorbed lipase, the encapsulated lipase shows higher thermal stability (4.9 times) and higher resistance to denaturants (5.0 times). Encouragingly, the encapsulated lipase shows high activity and reusability in lipase-catalyzed synthesis of propyl laurate, suggesting the potential application value of encapsulated lipase.


Assuntos
Enzimas Imobilizadas , Lipase , Lipase/metabolismo , Enzimas Imobilizadas/metabolismo , Microesferas , Adsorção , Estabilidade Enzimática
3.
Chem Biodivers ; 19(2): e202100675, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34866324

RESUMO

Hyperlipidemia (HLP) is a complex pathological condition results from lipid metabolism disorder, which is closely related to obesity, atherosclerosis and steatohepatitis. Emodin (EM), a natural anthraquinone, exhibits prominent hypolipidemic effects. However, its exact mechanism is still unclear. In this study, we successfully established hyperlipidemic zebrafish model induced by 4 % high-cholesterol diet (HCD) for 10 days and explored the anti-hyperlipidemic roles and underlying mechanisms of EM. The results indicated that EM attenuated the mortality and body mass index (BMI) of zebrafish with HLP, and ameliorated abnormal lipid levels involved in TC, TG, LDL-C and HDL-C levels. Besides, EM effectively reduced lipid accumulation in blood vessels and liver, alleviated hepatic histological damage, and inhibited vascular neutrophil inflammation. Finally, the mRNA expression of molecules related to lipid metabolism were studied by using real-time quantitative polymerase chain reaction (RT-qPCR) to investigated the underlying mechanism. Further results found that treatment with EM up-regulated AMPKα, LDLR, ABCA1 and ABCG1, and down-regulated SREBP-2, PCSK9 and HMGCR expression. In conclusion, EM showed a prominent mitigative effect on lipid metabolism disorder in zebrafish larvae with HCD-stimulated HLP, which was associated with the enhancement of LDL-C uptake and reverse cholesterol transport, and inhibition of cholesterol synthesis.


Assuntos
Emodina , Hiperlipidemias , Animais , Colesterol/metabolismo , Dieta Hiperlipídica/efeitos adversos , Emodina/farmacologia , Hiperlipidemias/tratamento farmacológico , Larva/metabolismo , Metabolismo dos Lipídeos , Fígado , Pró-Proteína Convertase 9/metabolismo , Pró-Proteína Convertase 9/farmacologia , Peixe-Zebra/metabolismo
4.
Front Pharmacol ; 12: 756924, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34621179

RESUMO

Liver fibrosis is a chronic pathological process that various pathogenic factors lead to abnormal hyperplasia of hepatic connective tissue, and its main feature is the excessive deposition of extracellular matrix. However, there are currently no drugs approved for the treatment of liver fibrosis. Phillygenin (PHI), a lignan isolated from Forsythiae Fructus, showed potential anti-inflammatory and anti-fibrosis effects but the mechanisms remain unknown. In view of the vital role of gut microbiota in the development of liver fibrosis, this study aimed to explore whether PHI could protect intestinal epithelial barrier and attenuate liver fibrosis by maintaining the homeostasis of intestinal microbiota. Therefore, the liver fibrosis model was induced by intraperitoneal injection of olive oil containing 10% carbon tetrachloride (CCl4) for 4 weeks in C57BL/6J mice. Histological analysis including Hematoxylin-Eosin, Masson, Sirius red, and immunohistochemistry staining were carried out to detect the histopathology and collagen deposition of mice liver tissues. The biochemical indexes related to liver function (ALT, AST, AKP, γ-GT), fibrosis (HYP, HAase, LN, PC III, IV-C) and inflammation (TNF-α, MIP-1, LPS) were determined by specific commercial assay kits. In vivo experimental results showed that PHI could improve liver histopathological injury, abnormal liver function, collagen deposition, inflammation and fibrosis caused by CCl4. Moreover, PHI restored the intestinal epithelial barrier by promoting the expression of intestinal barrier markers, including ZO-1, Occludin and Claudin-1. More importantly, the corrective effect of PHI on the imbalance of gut microbiota was confirmed by sequencing of the 16 S rRNA gene. In particular, PHI treatment enriches the relative abundance of Lactobacillus, which is reported to alleviate inflammation and fibrosis of damaged liver. Collectively, PHI attenuates CCl4-induced liver fibrosis partly via modulating inflammation and gut microbiota.

5.
Artigo em Inglês | MEDLINE | ID: mdl-33034915

RESUMO

To establish cholyglycine (CG) detection via enzyme multiplied immunoassay technique (EMIT), glucose-6-phosphate dehydrogenase (G6PD) was used as a reporter enzyme to prepare hapten-enzyme conjugate. Gel electrophoresis and UV scanning demonstrated that G6PD was successfully labeled with cholyglycine and CG-G6PD conjugate was obtained. Furthermore, the effects of various parameters on the preparation of CG-G6PD conjugates were investigated. Consequently, CG amount, NADH, D-glucose-6-phosphate (G6P), phosphate buffer and the pH, and ionic strength of solution had important effects on the residual activity of CG-G6PD. Moreover, CG amount, the pH, and G6P played important roles in changing CG labeling location on G6PD. Using the CG-G6PD conjugate as test kit, the cholyglycine-EMIT calibration curve was established, which could be employed in clinical detection of cholyglycine. This study provides some valuable information for preparing hapten-G6PD conjugates. This article is protected by copyright. All rights reserved.

6.
Biotechnol Appl Biochem ; 67(2): 257-264, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31651049

RESUMO

To establish cholyglycine (CG) detection via enzyme-multiplied immunoassay technique (EMIT), glucose-6-phosphate dehydrogenase (G6PD) was used as a reporter enzyme to prepare hapten-enzyme conjugate. Gel electrophoresis and UV scanning demonstrated that G6PD was successfully labeled with cholyglycine, and CG-G6PD conjugate was obtained. Furthermore, the effects of various parameters on the preparation of CG-G6PD conjugates were investigated. Consequently, CG amount, nicotinamide adenine dinucleotide, d-glucose-6-phosphate (G6P), phosphate buffer and the pH, and ionic strength of solution had important effects on the residual activity of CG-G6PD. Moreover, CG amount, the pH, and G6P played important roles in changing CG labeling location on G6PD. Using the CG-G6PD conjugate as test kit, the cholyglycine-EMIT calibration curve was established, which could be employed in clinical detection of cholyglycine. This study provides some valuable information for preparing hapten-G6PD conjugates.


Assuntos
Glucosefosfato Desidrogenase/metabolismo , Ácido Glicocólico/análise , Ácido Glicocólico/metabolismo , Leuconostoc mesenteroides/enzimologia
7.
Cancer Cell Int ; 18: 87, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29977159

RESUMO

BACKGROUND: Semaphorin 5A has been linked to tumor growth, invasion, and metastasis in pancreatic cancer. However, the role of semaphorin 5A in cervical cancer is not known. Our aim is to investigate the prognostic value of semaphorin 5A and its potential role in lymphangiogenesis and invasion in cervical cancer. METHODS: In this study, pathological features and clinical data of 232 cervical cancer patients were retrospectively reviewed. Semaphorin 5A protein and mRNA expression was detected by immunohistochemistry and quantitative real-time reverse transcription-polymerase chain reaction, respectively. In vitro, we determined the role and mechanistic pathways of semaphorin 5A in tumor progression in cervical carcinoma cell lines. RESULTS: Semaphorin 5A expression was significantly higher in stage IIb tumors than in stage Ia, Ib, and IIa tumors. High semaphorin 5A expression was significantly associated with pelvic lymph node metastasis, lymphovascular permeation, and poor survival. Semaphorin 5A induced lymphangiogenesis through a plexin-B/Met/vascular endothelial growth factor-C pathway. Semaphorin 5A also increased cervical cancer cell invasion by stimulating the expression and activity of matrix metalloproteinase-2 and matrix metalloproteinase-9 via PI3K/AKT and plexin-B3. CONCLUSION: Our findings indicate that semaphorin 5A may represent a poor prognostic biomarker and anti-metastasis therapeutic target in cervical cancer.

8.
Nanoscale ; 8(32): 15071-8, 2016 Aug 11.
Artigo em Inglês | MEDLINE | ID: mdl-27481652

RESUMO

Plasmon-mediated energy transfer is highly desirable in photo-electronic nanodevices, but the direct injection efficiency of "hot electrons" in plasmonic photo-detectors and plasmon-sensitized solar cells (plasmon-SSCs) is poor. On another front, Fano resonance induced by strong plasmon-exciton coupling provides an efficient channel of coherent energy transfer from metallic plasmons to molecular excitons, and organic dye molecules have a much better injection efficiency in exciton-SSCs than "hot electrons". Here, we investigate enhanced light-harvesting of chlorophyll-a molecules strongly coupled to Au nanostructured films via Fano resonance. The enhanced local field and plasmon resonance energy transfer are experimentally revealed by monitoring the ultrafast dynamical processes of the plexcitons and the photocurrent flows of the assembled plexciton-SSCs. By tuning the Fano factor and anti-resonance wavelengths, we find that the local field is largely enhanced and the efficiency of plexciton-SSCs consisting of ultrathin TiO2 films is significantly improved. Most strikingly, the output power of the plexciton-SSCs is much larger than the sum of those of the individual plasmon- and exciton-SSCs. Our observations provide a practical approach to monitor energy and electron transfer in plasmon-exciton hybrids at a strong coupling regime and also offer a new strategy to design photovoltaic nanodevices.

9.
Anat Rec (Hoboken) ; 299(6): 741-7, 2016 06.
Artigo em Inglês | MEDLINE | ID: mdl-26917449

RESUMO

Vascular endothelial growth factor-C (VEGF-C) is a key regulator in lymphangiogenesis, and is overexpressed in various malignancies. Integrin α4ß1, a new member of the VEGF-C/VEGF receptor pathway, was found to be overexpressed in melanoma tumors. However, little is known regarding the potential role of integrin α4ß1 in lymphangiogenesis and other solid tumors. The aim of this study was to investigate the expression patterns of integrin α4 and VEGF-C in relation to lymphangiogenesis and clinicopathological parameters in human colon cancer. The expression of integrin α4, VEGF-C, and VEGFR-3 was assessed in 71 human colon cancer tissues and 30 paracancerous normal tissues by immunohistochemical staining. Lymphatic microvessel density (LMVD) was measured after D2-40-labeling, and the correlations among different factors were statistically analyzed. The expression of integrin α4, VEGF-C, VEGFR-3, and LMVD was higher in colon cancer tissues compared with the normal paracancerous colon tissues. There was a positive correlation between the expression of integrin α4 and VEGF-C. Integrin α4 and VEGF-C were significantly associated with the clinicopathological parameters (LMVD, Duke's stage, and lymph node metastasis). Kaplan-Meier analyses indicated that patients with high integrin α4 or VEGF-C expression had significantly shorter overall survival and tumor-free survival time. Multivariate analyses suggested that integrin α4 and VEGF-C may serve as independent prognostic factors for human colon cancer. Both integrin α4 and VEGF-C are involved in lymphangiogenesis and lymphatic metastasis. Our results demonstrated that integrin α4 is a novel prognostic indicator for human colon cancer. Anat Rec, 299:741-747, 2016. © 2016 Wiley Periodicals, Inc.


Assuntos
Neoplasias do Colo/patologia , Integrina alfa4/metabolismo , Linfangiogênese/fisiologia , Metástase Linfática/patologia , Fator C de Crescimento do Endotélio Vascular/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias do Colo/metabolismo , Neoplasias do Colo/mortalidade , Feminino , Humanos , Vasos Linfáticos/metabolismo , Vasos Linfáticos/patologia , Masculino , Microvasos/metabolismo , Microvasos/patologia , Pessoa de Meia-Idade , Prognóstico , Taxa de Sobrevida , Regulação para Cima , Receptor 3 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Adulto Jovem
10.
Nanoscale ; 7(18): 8503-9, 2015 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-25896476

RESUMO

Strong coupling of plasmons and molecules generates intriguingly hybridized resonance. The IR-806 molecule is a near-infrared cyanine liquid crystal dye with multiple molecular bands and its tunable absorption spectrum varies dramatically with concentration. In this article, we investigate multiple hybridized resonances of the Au nanorods (AuNRs) strongly coupled to IR-806 molecules. Five hybridized resonance peaks are observed in the extinction spectra of the AuNR@IR-806 hybrids. Two resonance peaks at approximately 840 and 912 nm in the hybrids are reported for the first time. The dependence of the multiple hybridized peaks on the bare plasmon resonance wavelength of AuNRs and the molecular concentration is also demonstrated. The observations presented herein provide a plasmon-molecule coupling route for tuning optical responses of liquid crystal molecules.

11.
Nano Lett ; 15(4): 2705-10, 2015 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-25756956

RESUMO

Recent studies of the coupling between the plasmonic excitations of metallic nanostructures with the excitonic excitations of molecular species have revealed a rich variety of emergent phenomena known as plexcitonics. Here, we use a combined experimental and theoretical approach to demonstrate new and intriguing aspects in the ultrafast nonlinear responses of strongly coupled hybrid Fano systems consisting of gold nanorods decorated with near-infrared dye molecules. We show that the severely suppressed linear absorption around the Fano dip significantly enhances the unidirectional energy transfer from the plasmons to the excitons and further allows one-photon nonlinearity to be drastically and reversibly tuned. These striking observations are interpreted within a microscopic model stressing on two competing processes: saturated plasmonic absorption and weakened destructive Fano interference from the bleached excitonic absorption. The unusually strong one-photon nonlinearity revealed here provides a promising strategy in fabricating nanoplasmonic devices with both pronounced nonlinearities and good figures of merit.

12.
Cell Transplant ; 22(6): 1011-21, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23043768

RESUMO

Angiogenesis is a major obstacle for wound healing in patients with diabetic foot wounds. Mesenchymal stem cells (MSCs) have an important function in wound repair, and neurotrophin-3 (NT-3) can promote nerve regeneration and angiogenesis. We investigated the effect of NT-3 on accelerating wound healing in the diabetic foot by improving human bone marrow MSC (hMSC) activation. In vitro, NT-3 significantly promoted VEGF, NGF, and BDNF secretion in hMSCs. NT-3 improved activation of the hMSC conditioned medium, promoted human umbilical vein endothelial cell (HUVEC) proliferation and migration, and significantly improved the closure rate of HUVEC scratches. In addition, we produced nanofiber mesh biological tissue materials through the electrospinning technique using polylactic acid, mixed silk, and collagen. The hMSCs stimulated by NT-3 were implanted into the material. Compared with the control group, the NT-3-stimulated hMSCs in the biological tissue material significantly promoted angiogenesis in the feet of diabetic C57BL/6J mice and accelerated diabetic foot wound healing. These results suggest that NT-3 significantly promotes hMSC secretion of VEGF, NGF, and other vasoactive factors and that it accelerates wound healing by inducing angiogenesis through improved activation of vascular endothelial cells. The hMSCs stimulated by NT-3 can produce materials that accelerate wound healing in the diabetic foot and other ischemic ulcers.


Assuntos
Diabetes Mellitus Experimental/patologia , Diabetes Mellitus Experimental/terapia , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/metabolismo , Neurotrofina 3/farmacologia , Comunicação Parácrina/efeitos dos fármacos , Cicatrização/efeitos dos fármacos , Adolescente , Adulto , Animais , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Meios de Cultivo Condicionados/farmacologia , Pé Diabético/patologia , Células Endoteliais da Veia Umbilical Humana/citologia , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Humanos , Masculino , Células-Tronco Mesenquimais/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Fator de Crescimento Neural/metabolismo , Regeneração/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/metabolismo , Adulto Jovem
13.
Plant Physiol ; 157(4): 1884-99, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22010108

RESUMO

Thioredoxins (Trxs) are a multigenic family of proteins in plants that play a critical role in redox balance regulation through thiol-disulfide exchange reactions. There are 10 members of the h-type Trxs in rice (Oryza sativa), and none of them has been clearly characterized. Here, we demonstrate that OsTRXh1, a subgroup I h-type Trx in rice, possesses reduction activity in vitro and complements the hydrogen peroxide sensitivity of Trx-deficient yeast mutants. OsTRXh1 is ubiquitously expressed in rice, and its expression is induced by salt and abscisic acid treatments. Intriguingly, OsTRXh1 is secreted into the extracellular space, and salt stress in the apoplast of rice induces its expression at the protein level. The knockdown of OsTRXh1 results in dwarf plants with fewer tillers, whereas the overexpression of OsTRXh1 leads to a salt-sensitive phenotype in rice. In addition, both the knockdown and overexpression of OsTRXh1 decrease abscisic acid sensitivity during seed germination and seedling growth. We also analyzed the levels of hydrogen peroxide produced in transgenic plants, and the results show that more hydrogen peroxide is produced in the extracellular space of OsTRXh1 knockdown plants than in wild-type plants, whereas the OsTRXh1 overexpression plants produce less hydrogen peroxide under salt stress. These results show that OsTRXh1 regulates the redox state of the apoplast and influences plant development and stress responses.


Assuntos
Ácido Abscísico/farmacologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Oryza/fisiologia , Reguladores de Crescimento de Plantas/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Tiorredoxina h/metabolismo , Sequência de Aminoácidos , Animais , Espaço Extracelular/metabolismo , Peróxido de Hidrogênio/metabolismo , Dados de Sequência Molecular , Oryza/efeitos dos fármacos , Oryza/genética , Oryza/crescimento & desenvolvimento , Oxirredução , Fenótipo , Filogenia , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/genética , Folhas de Planta/fisiologia , Folhas de Planta/ultraestrutura , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Coelhos , Espécies Reativas de Oxigênio/análise , Proteínas Recombinantes de Fusão , Plântula/efeitos dos fármacos , Plântula/genética , Plântula/fisiologia , Plântula/ultraestrutura , Alinhamento de Sequência , Estresse Fisiológico , Tiorredoxina h/genética
14.
Colloids Surf B Biointerfaces ; 82(2): 302-6, 2011 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-20888194

RESUMO

Core-shell magnetic poly(styrene-acrylamide-acrylic acid) microspheres with carboxyl groups were successfully synthesized via dispersion copolymerization in the presence of nano-particle of Fe(3)O(4). The microspheres were characterized by FTIR spectra. They were used as carrier to immobilize bovine serum albumin (BSA). To investigate the effect of the microsphere surface properties on the immobilization of BSA, a series of microspheres with different hydrophobic/hydrophilic surface characteristics were prepared by adjusting molar percentages of monomers. The results showed that microspheres with different hydrophobicities/hydrophilicities had different immobilized ratios of BSA. In comparison with microspheres having hydrophilic characteristics those with hydrophobic characteristics had a much higher immobilized ratio. The possible reasons for these observations are discussed. In addition, ester activation and coupling times were optimized with respect to immobilized ratio.


Assuntos
Magnetismo , Microesferas , Poliestirenos/química , Soroalbumina Bovina/química , Acrilatos/química , Animais , Bovinos , Ésteres , Óxido Ferroso-Férrico/química , Interações Hidrofóbicas e Hidrofílicas , Microscopia Eletrônica de Transmissão/métodos , Polímeros/química , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Propriedades de Superfície
15.
Yi Chuan ; 29(7): 844-50, 2007 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-17646151

RESUMO

T-DNA tagging method is a high throughput system for identifying and cloning novel genes from T-DNA-inserted mutant population created via genetic transformation by Agrobacterium tumefaciens. However, the efficiency of using T-DNA-inserted mutant population to clone genes in rice was much lower than in Arabidopsis. In this study, a rice tagged line with two copies of T-DNA segments inserted independently to each other was screened out via a series of verification tests, including the co-segregated analysis between the mutated character and the sequence of T-DNA or the genomic sequence flanking inserted T-DNA. From this tagged line, two inserted incidents were separated from the progeny population of a plant heterozygous in two tagged sites, and some plants with the target trait and one of the inserted incidents were obtained, which were important basic materials for the subsequently co-segregated analysis between the mutated character and the sequence of inserted T-DNA, and for cloning the mutant gene in future. Based on this study, we have some thoughts about the gene cloning from the T-DNA tagged lines with more than one inserted sequence independently and put forward to discuss with colleagues.


Assuntos
DNA Bacteriano/genética , Mutagênese Insercional , Oryza/genética , Oryza/crescimento & desenvolvimento , Fenótipo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Transformação Genética
16.
Yi Chuan Xue Bao ; 33(4): 319-29, 2006 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-16625830

RESUMO

T-DNA tagging technique has provided a powerful strategy for identifying new functional genes in plants, and the key for success is the discovery of T-DNA-inserted mutants with changed phenotype. In this study, we screened 4,416 rice T1 tagged lines generated by enhancer trap system integrated with GLL4/VP16-UAS elements from two transformed parents, ZH11 and ZH15. We found many lines showed obvious morphological mutations, including two types--fake-homozygous mutation and separating mutation. The mutation phenotype was related to 14 kinds of trait such as plant height, heading date, leaf shape, leaf color, tiller number, panicle shape, spikelet number, grain shape, disease-like mutation, male sterility, awn, and so on. Among them, plant height, heading date, leaf color and male sterility had a comparatively high mutation frequency (over 1%). The mutation frequency of plant height and leaf color had no significant change between different years or transformed parents, but the frequency of heading date and male sterility varied greatly, suggesting that environment had a great effect on the expression of latter two traits. By conducting continuously co-segregating analyses in T1 and T2 generation, we identified 3 T-DNA-inserted mutants with malformed panicle or spikelets, which would provide a base for cloning correlative functional genes. At the same time, we selected randomly 42 lines with mutation phenotype and obtained 40 flanking sequences from 39 tagged lines by plasmid rescue or TAIL-PCR, of which, 26 were vector backbone sequence, 14 had good identity to rice genome sequence. The BlastN result showed the T-DNA preferentially integrated into protein-coding region in plants.


Assuntos
DNA Bacteriano/genética , Mutagênese Insercional , Oryza/genética , Plantas Geneticamente Modificadas , DNA de Plantas/genética , Ecossistema , Mutação , Oryza/anatomia & histologia , Oryza/metabolismo , Fenótipo , Reação em Cadeia da Polimerase , Locos de Características Quantitativas , Sequências Repetidas Terminais
17.
Yi Chuan Xue Bao ; 32(5): 501-6, 2005 May.
Artigo em Chinês | MEDLINE | ID: mdl-16018261

RESUMO

A clonally propagated F2 population (F2CP), derived from the rice cross of Qimiaoxiang/91SP068, was used to map rolled leaf QTLs. As the parent Qimiaoxiang is an unrolling leaf variety, while 91SP068 is a medium rolling variety with about 34% rolling index. One major QTL, rl8, which came from 91SP068, was mapped between two flanking SSR markers, RM6954 and RM6841, on chromosome 5, with genetic distance 3.8 cM, and was 1.0 cM away from RM6954. Its additive effect estimated by composite interval mapping (CIM) was 9.61 in 2002 and 6.23 in 2003, and the dominance effect was also different between two years, -1.19 in 2002 and -4.44 in 2003, respectively. It explained about 20% - 33% of the total phenotypic variation between two years. Furthermore, an integrated physical and genetic map encompassing rl8 region was constructed, and the physical distance of the interval was 542 kb, and the ratio of physical to genetic distance was 144 kb/cM. Based on this research, fine mapping of rolled leaf QTLs will not only facilitate the map-based cloning of the gene itself, but also improve the efficiency of marker-assisted selection in plant breeding.


Assuntos
Cromossomos de Plantas/genética , Genes de Plantas , Oryza/genética , Mapeamento Físico do Cromossomo/métodos , Locos de Características Quantitativas , Cruzamentos Genéticos , Repetições de Microssatélites , Folhas de Planta/genética
18.
Yi Chuan Xue Bao ; 32(4): 399-405, 2005 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-16011032

RESUMO

We constructed an F2 clonal population of intercross,Teqing/Lemont, and identified two quantitative trait loci (QTLs) contributing to rice sheath blight resistance on chromosome 9 and 11. The two QTLs were qSB-9 and qSB-11, respectively. From the population, three clonal lines were selected by markers' band types on both sides of these two QTLs, qSB-9 and qSB-11. Two were double-susceptible parent with homozygous susceptible alleles of these two loci,and the other was named as double-resistant parent,of which these two loci were all homozygous resistant alleles. These parents were separately backcrossed to recurrent parents, Teqing or Lemont. From BC2F1, marker-assisted selection was conducted in each proceeding generation and all back-crossed plants in BC2F1 and BC4F1 were inoculated by short toothpicks incubated with a strain, RH-9 of the fungus for identification of the resistance. Results suggested that these two QTLs were selected effectively in each backcross generation and their positions were also verified in identification of resistance to rice sheath blight. In seedling nursery of BC3F2 population, plants were selected through marker-assisted selection, and were separately mixed as homozygous lines of double-susceptible alleles on the background of Teqing, double-susceptible and double-resistant on the background of Lemont. The homozygous lines and their recurrent parents were simultaneously planted on experiment fields of Agriculture Collage of Yangzhou University and Lixiahe District Institute of Agricultural Science. The inoculation was performed by a random-block test with two replicates at each site. The results indicated that 1) The difference of sheath blight disease development was highly significant among materials under the same genetic background,and the order of disease seriousness among different homozygous lines were: double-susceptible line on the background of Lemont > double-susceptible line on the background of Teqing > Lemont > Teqing > double-resistant line on the background of Lemont; 2) When the resistant allele of qSB-9 or qSB-11 solely existed in a plant, its disease rating was reduced about 1.2 score, and 2.0 score when they simultaneously existed on the background of Lemont; 3) No significant interaction between the two QTLs controlling sheath blight resistance and environments was found. These studies have laid a strong groundwork in operation and application, of these QTLs contributing to rice sheath blight resistance.


Assuntos
Genes de Plantas , Oryza/genética , Doenças das Plantas/genética , Característica Quantitativa Herdável , Alelos , Cromossomos de Plantas , Cruzamentos Genéticos , Marcadores Genéticos/genética , Genótipo , Imunidade Inata/genética , Oryza/classificação , Oryza/microbiologia , Doenças das Plantas/microbiologia , Seleção Genética
19.
Ai Zheng ; 24(6): 699-703, 2005 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-15946482

RESUMO

BACKGROUND & OBJECTIVE: Most of the studies on lymphatic metastasis mechanism of carcinoma are confined to distribution of lymphatics. This research was to observe distribution and morphologic features of the lymphatics in periphery region of carcinoma, and morphologic changes of lymphatic endothelia. METHODS: Specimens were obtained from 10 patients with gastric carcinoma and 10 patients with colon carcinoma; 20 mice models bearing colon carcinoma were established. Morphology of lymphatics and ultrastructure of lymphatic endothelia were observed under microscope. Number density and volume density of lymphatics in periphery region of carcinoma and normal region were measured using computer image analysis system; open rate and destructive rate of lymphatics were calculated. RESULTS: The lymphatics in periphery region of carcinoma were dilated; their walls were disintegrated. Lymphatic endothelia were dissolved and destroyed into broken fragments; the organellae showed pathologic changes. Number density and volume density of lymphatics were significantly higher in periphery region of colon carcinoma than in normal region [(10.2+/-1.7)/mm(2) vs. (5.1+/-0.8)/mm(2), P < 0.05û (1.5+/-0.2)% vs. (0.7+/-0.0)%, P < 0.05], and were significantly higher in periphery region of gastric carcinoma than in normal region [(8.0+/-0.9)/mm(2) vs. (3.4+/-0.6)/mm(2), P < 0.01; (1.6+/-0.3)% vs. (0.8+/-0.2)%, P < 0.05]. Open rate of lymphatics was significantly higher in periphery regions of mice model colon carcinoma, human gastric carcinoma, and colon carcinoma than in relevant normal regions (22.2% vs. 7.8%, 35.0% vs. 8.0%, and 25.8% vs. 5.0%, P < 0.05). Destructive rate of lymphatics was significantly higher in periphery regions of mice model colon carcinoma, and human gastric carcinoma than in relevant normal regions (20.1% vs. 0, and 35.3% vs. 2.0%, P < 0.05). CONCLUSION: Compare with the lymphatics in normal tissue, the lymphatics in periphery region of carcinoma tissue are dilated with disintegrated walls; the lymphatic endothelia are destroyed; the density of the lymphatics is increased.


Assuntos
Neoplasias do Colo/patologia , Endotélio Linfático/ultraestrutura , Vasos Linfáticos/patologia , Neoplasias Gástricas/patologia , Animais , Neoplasias do Colo/ultraestrutura , Feminino , Humanos , Masculino , Camundongos , Pessoa de Meia-Idade , Neoplasias Gástricas/ultraestrutura
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