RESUMEN
@#Objective To establish a patient-derived tumor xenograft (PDTX) model and to observe the latency and rate of tumor formation, tumor size, tumor invasion and metastasis of transplanted tumors. Methods Seven patients with chest tumor in Drum Tower Hospital from April to December 2015 were chosen. There were 5 males and 2 females with age ranging from 61-71 years, including 4 patients of esophageal tumor and 3 patients of lung tumor. PDTX model was established by surgical removal of fresh tumor tissues of these patients and transplantation in NODPrkdcem26Il2rgem26Nju subcutaneous (NCG) mice. The latency and rate of tumor formation, tumor size, tumor invasion and metastasis of transplanted tumors were observed, and pathology of HE staining and immunohistochemical testing results were compared between PDTX model and the patients. Results PDTX model was successfully established in 4 patients, and the success rate was 66.7%, including 2 patients of esophageal cancer. The PDTX model retained the differentiation, morphological and structural characteristics of original tumors. Conclusion Pathology and molecular biology characteristics of PDTX model are consistent with the original tumor, which can be an “avatar” of tumor patients for clinical pharmacodynamics screening and new drug research and development.
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The objective of the present study was to investigate the optimal nutritional requirements for mycelial growth and sporulation of entomopathogenic fungus Aschersonia aleyrodis Webber by orthogonal layout methods. Herein the order of effects of nutrient components on mycelial growth was tryptone > Ca2+ > soluble starch > folacin, corresponding to the following optimal concentrations: 1.58 percent Soluble Starch, 3.16 percent Tryptone, 0.2 mmol l-1 Ca2+ and 0.005 percent Folacin. The optimal concentration of each factors for sporulation was 1.16 percent lactose, 0.394 percent tryptone, 0.4 mmol l-1 Fe2+ and 0.00125 percent V B1, and the effects of medium components on sporulation were found to be in the order lactose > V B1 > Fe2+ > tryptone. Under the optimal culture conditions, the maximum production of mycelial growth achieved 20.05 g l-1 after 7 days of fermentation, while the maximum spore yield reached 5.23 ×10(10) spores l-1 after 22 days of cultivation. This is the first report on optimization of nutritional requirements and design of simplified semi-synthetic media for mycelial growth and sporulation of A. aleyrodis.
O objetivo deste estudo foi investigar as exigências nutricionais ótimas para o crescimento micelial e esporulação do fungo entomopatogênico Aschersonia aleyrodis Webber. A ordem dos efeitos dos nutrientes na multiplicação micelial foi triptona>Ca2+>amido solúvel>folacina, com as seguintes concentrações ótimas: amido solúvel 1,58 por cento, tritona 3,16 por cento, Ca2+ 0,2mmol.l-1 e folacina 0,005 por cento. Para a esporulação, a concentração ótima de cada fator foi: lactose 1,16 por cento, triptona 0,394 por cento, Fe2+ 0,4mmol.l-1 e V B1 0,00125 por cento, na seguinte ordem: lactose> V B1> Fe2+>tritona. Em condições ótimas de cultura, a produção máxima de micélio foi 20,05g.l-1 após 7 dias de fermentação, enquanto o rendimento máximo de esporos foi 5,23 x 10(10) esporos.l-1 após 22 dias de cultivo. Esse é o primeiro relato sobre otimização das exigências nutricionais e desenvolvimento de meio de cultura semi-sintético para crescimento micelial e esporulação de A. aleyrodis.
Asunto(s)
Medios de Cultivo , Fermentación , Hongos , Micelio/crecimiento & desarrollo , Esporas Fúngicas , Métodos , MétodosRESUMEN
Entomopathogenic fungus Verticillium lecanii is a promising whitefly and aphid control agent. Chitinases secreted by this insect pathogen have considerable importance in the biological control of some insect pests. An endochitinase gene Vlchit1 from the fungus was cloned and overexpressed in Escherichia coli. The Vlchit1 gene not only contains an open reading frame (ORF) which encodes a protein of 423 amino acids (aa), but also is interrupted by three short introns. A homology modelling of Vlchit1 protein showed that the chitinase Vlchit1 has a (α/β)8 TIM barrel structure. Overexpression test and Enzymatic activity assay indicated that the Vlchit1 is a functional enzyme that can hydrolyze the chitin substrate, so the Vlchit1 gene can service as a useful gene source for genetic manipulation leading to strain improvement of entomopathogenic fungi or constructing new transgenic plants with resistance to various fungal and insects pests.
O fungo entomopatogênico Verticillium lecanii é um agente promissor no controle da mosca-branca e do pulgão. As quitinases secretadas por esse patógeno de insetos têm uma grande importância no controle biológico de doenças causadas por insetos. Um gene de endoquitinase Vlchit1 desse fungo foi clonado e expresso em Escherichia coli. O gene Vlchit contém não apenas um ORF que codifica uma proteína de 423 aminoácidos, mas também é interrompido por três pequenos introns. A modelagem de homologia da proteína Vlchit1indicou que a quitinase Vlchit1 tem uma estrutura (α/β) 8 TIM barrel. Testes de expressão e de atividade enzimática indicaram que Vlchit1 é uma enzima funcional que hidroliza quitina, portanto o gene Vlchit pode ser um gene útil para manipulação genética para melhoramento de cepas de fungos entomopatogênicos ou para a construção de novas plantas transgênicas com resistência a várias doenças causadas por fungos e insetos.