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BACKGROUND: Formin, a highly conserved multi-domain protein, interacts with microfilaments and microtubules. Although specifically expressed formin genes in anthers are potentially significant in research on male sterility and hybrid wheat breeding, similar reports in wheat, especially in thermo-sensitive genic male sterile (TGMS) wheat, remain elusive. RESULTS: Herein, we systematically characterized the formin genes in TGMS wheat line BS366 named TaFormins (TaFHs) and predicted their functions in inducing stress response. In total, 25 TaFH genes were uncovered, majorly localized in 2A, 2B, and 2D chromosomes. According to the neighbor-joining (NJ) method, all TaFH proteins from wheat and other plants clustered in 6 sub-groups (A-F). The modeled 3D structures of TaFH1-A/B, TaFH2-A/B, TaFH3-A/B and TaFH3-B/D were validated. And different numbers of stress and hormone-responsive regulatory elements in their 1500 base pair promoter regions were contained in the TaFH genes copies. TaFHs had specific temporal and spatial expression characteristics, whereby TaFH1, TaFH4, and TaFH5 were expressed highly in the stamen of BS366. Besides, the accumulation of TaFHs was remarkably lower in a low-temperature sterile condition (Nanyang) than fertile condition (Beijing), particularly at the early stamen development stage. The pollen cytoskeleton of BS366 was abnormal in the three stages under sterile and fertile environments. Furthermore, under different stress levels, TaFHs expression could be induced by drought, salt, abscisic acid (ABA), salicylic acid (SA), methyl jasmonate (MeJA), indole-3-acetic acid (IAA), polyethylene glycol (PEG), and low temperature. Some miRNAs, including miR167, miR1120, and miR172, interacts with TaFH genes; thus, we constructed an interaction network between microRNAs, TaFHs, phytohormone responses, and distribution of cytoskeleton to reveal the regulatory association between upstream genes of TaFH family members and sterile. CONCLUSIONS: Collectively, this comprehensive analysis provides novel insights into TaFHs and miRNA resources for wheat breeding. These findings are, therefore, valuable in understanding the mechanism of TGMS fertility conversion in wheat.
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Fitomejoramiento , Triticum , Citoesqueleto/metabolismo , Fertilidad/genética , Forminas , Regulación de la Expresión Génica de las Plantas , Microtúbulos/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Polen/genética , Polen/metabolismo , Triticum/genética , Triticum/metabolismoRESUMEN
The lack of stable and efficient techniques to synthesize high-quality large-area thin films is one of the major bottlenecks for the real-world application of the 2D transition metal dichalcogenides. In this work, the growth of molybdenum disulfide (MoS2 ) on sapphire substrates by sulfurizing the MoO3 film deposited by atomic layer deposition (ALD) is reported. The advantages of the ALD method can be well inherited, and the synthesized MoS2 films exhibit excellent layer controllability, wafer-scale uniformity, and homogeneity. MoS2 films with desired thickness can be obtained by varying MoO3 ALD cycles. The atomic force microscope and Raman measurements demonstrate that the ALD-based MoS2 has good uniformity. Clear Raman shift as a function of the film thickness is observed. Field-effect transistor devices are fabricated through a transfer-free and top-down process. High On/Off current ratio (≈104 ) and medium-level electron mobilities (≈0.76 cm2 V-1 s-1 for monolayer, and 5.9 cm2 V-1 s-1 for four-layer) are obtained. The work opens up an attractive approach to realize the application of wafer-scale 2D materials in integrated circuits and systems.
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A halogen-atom-transfer (XAT)-based method for carbonylazotization of pyrroles or indoles with aryldiazonium salts and polyhalomethanes via dual C(sp2)-H bond functionalization is described. Using aryldiazonium salts realizes carbonylation/azotization of pyrroles or indoles via polyhalomethyl-radical-mediated and electrophilic substitution, thus providing a green, efficient, and step-economy approach for synthesis of multifunctional pyrroles or indoles from the easily available substrates. Notably, this strategy relies on the use of aryldiazonium salts to extend the well-established iodine atom transfer to bromine or chlorine atom transfer.
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OBJECTIVE: To establish the integrated discrete multiple organ cell culture (IdMOC) system. METHODS: Rat primary cell of hepatocyte, nephrocyte, cardiomyocytes, alveolar macrophage, dermal fibroblasts were isolated by collagenase digestion, separation of bronchial lavage, two-step digestion method and cultured respectively, with monolayer culture. To establish the integrated discrete multiple organ cell culture (IdMOC) system, glass slides of five different cells were used to the same dish with 10% FBS DMEM medium cultured 7d, using MTT comparison primary cells cultured alone and cocultured when growth. RESULTS: Established rat hepatocytes, renal cell, cardiomyocyte, alveolar macrophages, dermal fibroblasts separation method was stable, cell separation survival rate was about 90.0%. Hepatocytes separation survival rate 90.3% ,renal cell separation survival rate 91.9%, cardiomyocyte separation survival rate 93.0% and beating rate indifference curve among 3d-15d, alveolar macrophages cell separation survival rate 90.8%, dermal fibroblasts cell separation survival rate 92.7%. Five primary cells multiple organ cells coculture showed cocultured cell growth proliferation well, cultured alone and cocultured cells growth curve basic coincide. CONCLUSION: Established rat multiple organ cell co-culture is successful.
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Técnicas de Cultivo de Célula/métodos , Animales , Células Epiteliales/citología , Hepatocitos/citología , Macrófagos Alveolares/citología , Miocitos Cardíacos/citología , Ratas , Ratas Sprague-DawleyRESUMEN
γδT cells are unconventional T lymphocytes that bridge innate and adaptive immunity. Based on the composition of T cell receptor and the cytokines produced, γδT cells can be divided into diverse subsets that may be present at different locations, including the liver, epithelial layer of the gut, the dermis and so on. Many of these cells perform specific functions in liver diseases, such as viral hepatitis, autoimmune liver diseases, non-alcoholic fatty liver disease, liver cirrhosis and liver cancers. In this review, we discuss the distribution, subsets, functions of γδT cells and the relationship between the microbiota and γδT cells in common hepatic diseases. As γδT cells have been used to cure hematological and solid tumors, we are interested in γδT cell-based immunotherapies to treat liver diseases.
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Microbioma Gastrointestinal/inmunología , Inmunoterapia/métodos , Linfocitos Intraepiteliales/inmunología , Hepatopatías/inmunología , Animales , Modelos Animales de Enfermedad , Humanos , Mucosa Intestinal/inmunología , Mucosa Intestinal/microbiología , Linfocitos Intraepiteliales/trasplante , Hígado/inmunología , Hígado/patología , Hepatopatías/microbiología , Hepatopatías/patología , Hepatopatías/terapiaRESUMEN
OBJECTIVE: To explore effects of alcohol on the expression of gial fibral acidic protein (GFAP) and S100 of astrocytes in brain. METHODS: Different dosages of alcohol (2,5 and 20 mmol/L) were administered to astrocytes of fetal rat brain in vitro. The expression of GFAP and S100 of astrocytes was detected by immunocytochemistry, respectively. RESULTS: With the dosage of alcohol increasing, the expression of GFAP and S100 was reduced and the number of astrocytes in high dosage group was observed to be decreased slightly. CONCLUSION: It suggested that alcohol could repress the expression of GFAP and S100 of astrocytes, which might be the main mechanism of developmental abnormalities of central nervous system induced by alcohol.
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Astrocitos/metabolismo , Encéfalo/metabolismo , Etanol/toxicidad , Proteína Ácida Fibrilar de la Glía/metabolismo , Proteínas S100/metabolismo , Animales , Encéfalo/efectos de los fármacos , Encéfalo/embriología , Femenino , Feto , Masculino , Embarazo , Ratas , Ratas Sprague-DawleyRESUMEN
OBJECTIVE: To provide the proof for establishing the high throughput screening method of the acute toxicity by comparing the 4 screening methods of the acute toxicity in vitro. METHODS: HepG2 cells, plated in 96-well plates, were treated with 47 compounds and we detected the cytotoxicity using 4 methods and calculated the IC50, IC45, IC40, IC35, IC30, and these values were compared with the LD50. RESULTS: The IC45 from the neutral red method has the best relativity with the LD50. CONCLUSION: Plating HepG2 cells in 96-well plates, the cytotoxicity be detected using the neutral red method and the IC45 was calculated. It is considered to provide the proof for establishing the high throughput screening method of the acute toxicity in vitro.
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Rojo Neutro , Pruebas de Toxicidad Aguda/métodos , Supervivencia Celular/efectos de los fármacos , Células Hep G2 , Humanos , Concentración 50 Inhibidora , Dosificación Letal MedianaRESUMEN
OBJECTIVE: To explore the estrogenic effects and disruptive mechanism of NP and BPA by reporter gene-based assays we developed. METHODS: pERE-Luc plamid was generated by inserting estrogen response element (ERE) fragment into MCS of pGL3-promoter vector. MCF7 cells were cotransfected with pERE-Luc and phRL-SV40 using Sofast transfection reagent. The cells then treated with 17beta-estradiol (E2), tamoxifen (Tam), nonylphenol(NP) and bisphenol A (BPA) and expression of the repoter gene in the cell lysates was assayed using Dual-Lucferase reporter assay system. RESULTS: The pERE-Luc plasmid was constructed. Luciferase activities of MCF7 cells transfected pERE-Luc showed dose-responed realitionship with E2. 1 x 10(-11) mol/L E2 could induce the expression of reporter gene and 1 x 10(-9) mol/L E2 resulted in the largest luciferase activity. E2 couldn't induce the luciferase activity without pERE-Luc. Tam is a complete antagonist, inhibited the E2-induced luciferase expression. NP induced the luciferase activity at concertrations > 1 x 10(-6) mol/L, BPA induced the luciferase activity at concertrations > 1 x 10(-6) mol/L. The estrogenic activity of NP was more than BPA. CONCLUSION: The assay we established is usful, NP and BPA showed estrogenic activities.
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Disruptores Endocrinos/toxicidad , Estrógenos no Esteroides/toxicidad , Genes Reporteros/efectos de los fármacos , Fenoles/toxicidad , Compuestos de Bencidrilo , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Línea Celular Tumoral , Estradiol/farmacología , Femenino , Genes Reporteros/genética , Humanos , Receptores de Estrógenos/efectos de los fármacos , Receptores de Estrógenos/genética , Receptores de Estrógenos/metabolismoRESUMEN
OBJECTIVE: To study the expression of sHsps in normal palate and cleft palate during mouse embryogenesis. METHODS: At GD10, gestational mice of the treatment and the control were administered with 80 mg/kg retinoic acid and the same volume vegetable oil separately, and the normal palate and cleft palate of embryos were harvested in GD15-GD17. The relative abundance of sHsps of all samples was measured by reverse transcript polymerase chain reaction (RT-PCR). RESULTS: In the normal limbs, except that there is no expression of Hspb10 at GD17, all the other Hsps expressed obviously in GD15-GD17. To the normal palates, the expressional abundance of Hsp20, Hsp25, Hsp27, Hsp32, Hspb2, Hspb3, Hspb7 was stable, that of Hsp30, Hspb5 was increased following the embryos aging, and the expressional peak of Hsp10, Hsp22, Hspb4 occurred at GD16. In GD15-GD17, the expressional abundance of Hsp30, Hsp32, Hspb4, Hspb10 of the cleft palates was higher than that of the normal palates, but the expressional abundance of Hsp60, Hspb5, Hspb9 of the cleft palates was lower than that of the normal palates. The expressional models of Hspb9, Hspb 10 of the normal palates were different from those of the cleft palates obviously. CONCLUSION: Except that there is no expression of HspblO at GD17, all the other Hsps expressed obviously in GD15-GD17 during normal clefts' development. To different Hsps, there is different expressional characteristic. Hsp30, Hsp32, Hspb4, Hspb10 maybe play a protective role in the stress action of cleft palate, and Hsp10, Hsp60, Hspb5, Hspb9, Hspb10 were maybe relative to cleft palate.
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Fisura del Paladar/embriología , Fisura del Paladar/genética , Proteínas de Choque Térmico/metabolismo , Animales , Fisura del Paladar/inducido químicamente , Fisura del Paladar/metabolismo , Embrión de Mamíferos/embriología , Embrión de Mamíferos/metabolismo , Femenino , Regulación del Desarrollo de la Expresión Génica , Proteínas de Choque Térmico/genética , Masculino , Ratones , Ratones Endogámicos ICR , TretinoinaRESUMEN
A new dihydrochalcone, 4'-hydroxy-4,2'-dimethoxy-dihydrochalcone, was isolated from Chinese dragon's blood, the red resin of Dracaena cochinchinensis. Its structure was established by spectrum analysis.
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Chalconas/aislamiento & purificación , Dracaena/química , Plantas Medicinales/química , Chalconas/química , Estructura Molecular , Resonancia Magnética Nuclear Biomolecular , Resinas de Plantas/químicaRESUMEN
OBJECTIVES: A total of 2402 patients with arsenic-related skin lesions, such as hyperkeratosis, hyperpigmentation or hypopigmentation, or even skin cancer in a few villages in Southwest Guizhou Autonomous Prefecture, China represent a unique case of endemic arsenism related with indoor combustion of high arsenic coal. This study aimed to investigate the cluster of arsenism cases and the possible relevant factors including GSTP1 polymorphism in two clans of different ethnic origin living in one village for generations. METHODS: A questionnaire-based study was performed in 170 Miao clan P members, 10 of whom had arsenic-related skin diseases, and 153 Han clan G1 members, 50 of whom had arsenic-related skin diseases. The data were checked against the registration archives since the 1980s. At the same time, arsenic concentrations in samples of coal, indoor air, drinking water, corn and chilli pepper that were once baked over the stoves for desiccation, as well as in samples of urine and hair of clan members were determined. Glutathione S-transferase P1 (GSTP1) A1578G polymorphism was genotyped by a restriction fragment length polymorphism-based procedure. RESULTS: Arsenism morbidity in Miao clan P was significantly lower than in the neighbouring Han clan G1 [5.9 vs. 32.7%, odds ratio (OR)=0.13, 95% confidence interval (CI): 0.06-0.27, P<0.0001]. No sex differences were confirmed inside both clans. Analyses of the environmental samples indicated that Miao clan P members were exposed to higher amounts of arsenic via inhalation and food ingestion. Hair and urine samples also proved a higher arsenic body burden in ethnic Miao individuals. No corresponding differences by sex were found. Higher frequencies of combined mutant genotype G/G1578 and A/G1578 (OR=4.72, 95% CI: 2.34-9.54, P<0.0001) and of mutant allele G1578 (OR=3.22, 95% CI: 2.00-5.18, P<0.0001) were detected in diagnosed arsenism patients than in non-diseased individuals. The Miao individuals showed a lower percentage of combined mutant genotypes (30.6 vs. 52.7%, OR=0.40, 95% CI: 0.19-0.84, P=0.015) as well as of mutant allele G1578 (OR=0.46, 95% CI: 0.24-0.88, P=0.017) than their Han neighbours. CONCLUSIONS: Genetic predisposition influences dermal arsenism toxicity. The GSTP1 A1578G (Ile105Val) status might be a susceptibility factor for arsenic-related skin lesions.