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1.
Opt Lett ; 49(9): 2445-2448, 2024 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-38691740

RESUMEN

Spatial-temporal optical vortices (STOVs) have recently become the focus of newly structured optical fields. In this paper, their propagation on a 2D curved surface named the constant Gaussian curvature surface (CGCS) is studied. Using the matrix optics approach, we provide the analytical solution of the STOV propagation under the paraxial approximation on the CGCS with positive curvature. One method of creating timers is made possible by the spatiotemporal distribution direction of STOV light intensity, which swings like a pendulum throughout the evolution, in contrast to propagation on a flat surface. This swing, however, stops when the curved surface's curvature radius matches the light's Rayleigh distance. Besides, the transverse orbital angular momentum of STOV is deduced, and we find that the intrinsic and extrinsic OAM periodically exchange, but the total transverse OAM is always zero during the propagation on CGCS. It aids in controlling the transverse extrinsic orbital angular momentum of STOV in nontrivial space.

2.
Explor Target Antitumor Ther ; 5(1): 34-53, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38464391

RESUMEN

Colorectal cancer (CRC) is a multifaceted disease influenced by the interplay of genetic and environmental factors. The clinical heterogeneity of CRC cannot be attributed exclusively to genetic diversity and environmental exposures, and epigenetic markers, especially DNA methylation, play a critical role as key molecular markers of cancer. This review compiles a comprehensive body of evidence underscoring the significant involvement of DNA methylation modifications in the pathogenesis of CRC. Moreover, this review explores the potential utility of DNA methylation in cancer diagnosis, prognostics, assessment of disease activity, and prediction of drug responses. Recognizing the impact of DNA methylation will enhance the ability to identify distinct CRC subtypes, paving the way for personalized treatment strategies and advancing precision medicine in the management of CRC.

3.
MycoKeys ; 100: 95-121, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-38025587

RESUMEN

The genus Ciliochorella is a group of pestalotioid fungi, which typically occurs in subtropical and tropical areas. Species from the Ciliochorella genus play important roles in the decomposition of litter. In this study, we introduce two new species (Ciliochorellachinensissp. nov. and C.savannicasp. nov.) that were found on leaf litter collected from savanna-like vegetation in hot dry valleys of southwestern China. Phylogenetic analyses of combined LSU, ITS and tub2 sequence datasets indicated that C.chinensis and C.savannica respectively form a distinct clade within the Ciliochorella genus. The comparison of the morphological characteristics indicated that the two new species are well differentiated within this genus species. Analysis of the evolutionary history suggests that Ciliochorella originated from the Eurasian continent during the Paleogene (38 Mya). Further, we find that both new species can produce cellulase and laccase, playing a decomposer role.

4.
AMB Express ; 13(1): 118, 2023 Oct 21.
Artículo en Inglés | MEDLINE | ID: mdl-37864620

RESUMEN

Telomere and telomerase are crucial factors in cell division and chromosome stability. Telomerase activity in most cells depends on the transcription control by the telomerase reverse transcriptase (TERT). The introduction of an exogenous human TERT (hTERT) in cultured cells could enhance telomerase activity and elongate the lifespan of various cells. Telomere elongation mechanisms vary between insects and are complex and unusual. Whether the use of exogenous hTERT can immortalize primary insect cells remains to be investigated. In this study, we used a recombinant virus expressing hTERT to infect primary cultured cells of Periplaneta americana and evaluated its effects on insect cell immortalization. We found that hTERT was successfully expressed and promoted the growth of P. americana cells, shortening their doubling time. This was due to the ability of hTERT to increase the activity of telomerase in P. americana cells, thus prolonging the telomeres. Our study lays the foundation for understanding the mechanisms of telomere elongation in P. americana, and suggests that the introduction of hTERT into insect cells could be an efficient way to establish certain insect cell lines.

6.
Pol J Microbiol ; 71(3): 301-307, 2022 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-36047456

RESUMEN

This study aims to evaluate associations between the immunochromatographic rapid test technique and Trichomonas vaginalis (TV) infection in patients with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) in Taiwan. All patients received post-prostate massage urine (VB3) Trichomonas rapid tests. The demographic characteristics and urogenital symptoms of CP/CPPS were recorded. Routine urinalysis of VB3 was also performed, and laboratory examination results of semen were recorded if available. A total of 29 patients with TV infection and 109 without TV infection were enrolled, which reflected that the prevalence in patients with TV infection was approximately 21%. Patients with TV infection displayed a significantly higher frequency of suprapubic/lower abdominal pain (p = 0.034), semen leukocyte > 5/high-power field (HPF) (p = 0.020), and an inflammatory type (category IIIA) (p = 0.005) than patients without TV infection. A higher prevalence of TV infection was found in patients with category IIIA (47.37%). No significant difference was found in the symptom duration and other clinical symptoms. In conclusion, the high prevalence of TV infection was revealed in CP/CPPS patients using the VB3 rapid Trichomonas test, especially in CP/CPPS patients with category IIIA. Thus, rapid TV testing might be vital for CP/CPPS patients in the hospital.


Asunto(s)
Prostatitis , Trichomonas vaginalis , Enfermedad Crónica , Humanos , Masculino , Dolor Pélvico/diagnóstico , Prostatitis/diagnóstico , Semen , Síndrome
7.
Genes (Basel) ; 13(8)2022 07 29.
Artículo en Inglés | MEDLINE | ID: mdl-36011275

RESUMEN

The white wax secreted by the male insects of the Chinese white wax scale (CWWS) is a natural high-molecular-weight compound with important economic value. However, its regulatory mechanism of wax biosynthesis is still unclear. In this study, a weighted gene coexpression network analysis (WGCNA) was used to analyze transcriptome data of first- and second-instar females, early and late female adults, and first- and second-instar males. A total of 19 partitioned modules with different topological overlaps were obtained, and three modules were identified as highly significant for wax secretion (p < 0.05). A total of 30 hub genes were obtained through screening, among which elongation of very-long-chain fatty acids protein (ELOVL) and fatty acyl-CoA reductase (FAR) are important catalytic enzymes of fatty acid metabolism. Furthermore, their metabolic catalytic products are involved in the synthesis of wax biosynthesis. The results demonstrate that WGCNA can be used for insect transcriptome analysis and effectively screen out the key genes related to wax biosynthesis.


Asunto(s)
Hemípteros , Animales , China , Femenino , Perfilación de la Expresión Génica , Hemípteros/genética , Masculino , Transcriptoma/genética
8.
In Vitro Cell Dev Biol Anim ; 58(5): 365-375, 2022 May.
Artículo en Inglés | MEDLINE | ID: mdl-35653076

RESUMEN

The baculovirus expression vector system using insect cells as a bioreactor has been used for in vitro expression of recombinant proteins and plays an important role in the fields of biology, agronomy, and medicine. Screening suitable host cell lines is an important part of the construction of insect cell baculovirus expression systems. In previous research, we used a single-cell cloning process with the Papilio xuthus cell line RIRI-PX1 and obtained the monoclonal cell line RIRI-PX1-C31. In this study, we compared the basic biological and recombinant protein expression characteristics of RIRI-PX1-C31 and its parent cell line RIRI-PX1 and found that the expression of recombinant ß-galactosidase in RIRI-PX1-C31 was significantly higher than that in the parental cell line. Further serum-free adaptation studies confirmed that RIRI-PX1-C31 can adapt to the growth environment of Express Five Serum-free medium and that its expression level of recombinant ß-galactosidase was significantly higher than that before adaptation.


Asunto(s)
Mariposas Diurnas , Animales , Baculoviridae/genética , Línea Celular , Células Clonales , Expresión Génica , Insectos , Proteínas Recombinantes/metabolismo , beta-Galactosidasa/genética , beta-Galactosidasa/metabolismo
9.
In Vitro Cell Dev Biol Anim ; 58(4): 278-288, 2022 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-35460045

RESUMEN

Although the baculovirus expression vector system (BEVS) is widely used in the production of recombinant proteins, only a few lepidopteran insect cell lines have been successfully used so far. This study aimed at evaluating the characteristics of an embryonic cell line from the American cockroach Periplaneta americana L., RIRI-PA1, and determining whether it could be used in recombinant protein expression. Wild type Autographa californica multiple nucleopolyhedrovirus (AcMNPV-wt) and green fluorescent protein (GFP)-replicating recombinant baculoviruses (AcMNPV-GFP) were used to infect RIRI-PA1 respectively, demonstrating that RIRI-PA1 cells could be infected by AcMNPV and express recombinant proteins. Within 24 h of infection with AcMNPV-GFP, the GFP expression was higher than that in Sf21 cells. Furthermore, the infection of RIRI-PA1 cells increased gradually (multiplicity of infection, 10) within 24 h, while in Sf21 cells, the infection only began to increase within 48 h. However, after exposure for 96-168 h, the virus progeny and recombinant protein production of RIRI-PA1 cells was lower than those of Sf21 cells. Western blotting revealed that RIRI-PA1 cells could express recombinant GFP, and the protein expression level positively correlated with the multiplicity of infection. In conclusion, this is the first report that a cell line from P. americana has shown susceptibility to infection from a baculovirus and likewise express recombinant protein. Although the yield of recombinant GFP was not as high as that of Sf21 cells, the results nonetheless showed that RIRI-PA1 had an infection rate advantage in the short term (within 24 h of infection), which is of great value for further development and utilization.


Asunto(s)
Cucarachas , Periplaneta , Animales , Baculoviridae , Línea Celular , Cucarachas/metabolismo , Proteínas Fluorescentes Verdes/metabolismo , Nucleopoliedrovirus , Proteínas Recombinantes/metabolismo , Spodoptera
10.
Front Cell Dev Biol ; 9: 759813, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34778269

RESUMEN

Background: Colorectal cancer (CRC) is a leading cause of cancer death, and early diagnosis of CRC could significantly reduce its mortality rate. Previous studies suggest that the DNA methylation status of zinc finger genes (ZFGs) could be of potential in CRC early diagnosis. However, the comprehensive evaluation of ZFGs in CRC is still lacking. Methods: We first collected 1,426 public samples on genome-wide DNA methylation, including 1,104 cases of CRC tumors, 54 adenomas, and 268 para-tumors. Next, the most differentially methylated ZFGs were identified and validated in two replication cohorts comprising 218 CRC patients. Finally, we compared the prediction capabilities between the ZFGs and the SEPT9 in all CRC patients and the KRAS + and KRAS- subgroup. Results: Five candidate ZFGs were selected: ESR1, ZNF132, ZNF229, ZNF542, and ZNF677. In particular, ESR1 [area under the curve (AUC) = 0.91] and ZNF132 (AUC = 0.93) showed equivalent or better diagnostic capability for CRC than SEPT9 (AUC = 0.91) in the validation dataset, suggesting that these two ZFGs might be of potential for CRC diagnosis in the future. Furthermore, we performed subgroup analysis and found a significantly higher diagnostic capability in KRAS + (AUC ranged from 0.97 to 1) than that in KRAS- patients (AUC ranged from 0.74 to 0.86) for all these five ZFGs, suggesting that these ZFGs could be ideal diagnostic markers for KRAS mutated CRC patients. Conclusion: The methylation profiles of the candidate ZFGs could be potential biomarkers for the early diagnosis of CRC, especially for patients carrying KRAS mutations.

11.
Opt Express ; 29(18): 28631-28642, 2021 Aug 30.
Artículo en Inglés | MEDLINE | ID: mdl-34614989

RESUMEN

The Rindler space-time describing a series of accelerating observers is Ricci flat, but it still has novel optical effects. In the case of Wenzel, Kramers, and Brillouin (WKB) approximation, we derive the light paths in the Rindler frame based on the covariant wave equation and geodesic equations. Then, we use ABCD matrix optics method to explore the propagation characteristics of Rindler frame, thus link three different optical transformation scenes (geometry, gravity, and vacuum refractive index) together. Moreover, the propagation characteristics of hollow beam in Rindler space-time are described analytically. In the longitudinal direction, we demonstrate the shift and stretch effects of the dark spot of a beam, while the transverse spot size is proved to be convergence in the accelerated system, and the wavefront curvature can tend a constant twice the acceleration at the far field. Those characteristics are quite different from the ones in the flat space-time. Based on these calculations, we simply demonstrate the position uncertain relationship between the transverse beam size and the momentum, which surprisingly coincides with the derivation of quantization. We hope that we can provide one simple method to analyze the beam propagation in the accelerated frame.

12.
Medicine (Baltimore) ; 100(33): e27007, 2021 Aug 20.
Artículo en Inglés | MEDLINE | ID: mdl-34414992

RESUMEN

ABSTRACT: The objective of this report was to introduce a new suture augmentation of coracoclavicular (CC) and acromioclavicular ligament reconstruction for acute Rockwood grade III to V acromioclavicular dislocations.From January 2015 to January 2019, 43 patients with Rockwood III to VI acute acromioclavicular dislocations were retrospectively reviewed. For comparison, another series of 28 patients treated with double Endobutton technique from January 2011 to December 2014 were reviewed. A P < .05 was considered statistical significance.The mean follow-up period of the 2 series were 39.69 ±â€Š7.42 months (range, 24-54 months) and 37.86 ±â€Š8.23 months (range, 26-48 months) (P > .05), respectively. There were significant differences regarding CC space (11.62 ±â€Š2.54 mm vs 16.78 ±â€Š5.53 mm; P < .05), CC reduction loss (5.56 ±â€Š4.73 mm vs 26.25 ±â€Š4.42 mm; P < .05), and acromioclavicular space (6.89 ±â€Š1.87 mm vs 7.95 ±â€Š2.37 mm; P < .05). There were significant differences regarding the disabilities of the arm, shoulder, and hand questionnaire (3.3 ±â€Š2.8 vs 5.32 ±â€Š4.37; P < .05) and University of California-Los Angeles shoulder rating scale (31.19 ±â€Š2.48 vs 29.24 ±â€Š2.48; P < .05). The excellent to good percentages were 100% (n = 32) and 85% (n = 23), respectively.In conclusion, the suture augmentation of acromioclavicular and CC ligament reconstruction is a reliable technique for acute acromioclavicular dislocation with minimal complications.Type of study/level of evidence: Therapeutic IIa.


Asunto(s)
Articulación Acromioclavicular/cirugía , Ligamentos/cirugía , Suturas/normas , Articulación Acromioclavicular/fisiopatología , Adulto , Femenino , Humanos , Ligamentos/fisiopatología , Masculino , Persona de Mediana Edad , Procedimientos Ortopédicos/métodos , Estudios Retrospectivos , Luxación del Hombro/cirugía , Suturas/efectos adversos , Resultado del Tratamiento
13.
Clin Epigenetics ; 13(1): 89, 2021 04 23.
Artículo en Inglés | MEDLINE | ID: mdl-33892791

RESUMEN

BACKGROUND: Aberrant DNA methylation has been firmly established as a factor contributing to the pathogenesis of colorectal cancer (CRC) via its capacity to silence tumour suppressor genes. However, the methylation status of multiple tumour suppressor genes and their roles in promoting CRC metastasis are not well characterised. METHODS: We explored the methylation and expression profiles of CPEB1 (the gene encoding cytoplasmic polyadenylation element-binding protein 1), a candidate CRC tumour suppressor gene, using The Cancer Genome Atlas (TCGA) database and validated these results in both CRC cell lines and cells from Han Chinese CRC patients (n = 104). The functional role of CPEB1 in CRC was examined in experiments performed in vitro and in vivo. A candidate transcription factor capable of regulating CPEB1 expression was predicted in silico and validated by luciferase reporter, DNA pull-down, and electrophoretic mobility shift assays. RESULTS: Hypermethylation and decreased expression of CPEB1 in CRC tumour tissues were revealed by TCGA database. We also identified a significant inverse correlation (Pearson's R = - 0.43, P < 0.001) between promoter methylation and CPEB1 expression. We validated these results in CRC samples and two CRC cell lines. We also demonstrated that up-regulation of CPEB1 resulted in significantly decreased tumour growth, migration, invasion, and tumorigenicity and promoted tumour cell apoptosis both in vitro and in vivo. We identified the transcription factors CCAAT enhancer-binding protein beta (CEBPB) and transcription factor CP2 (TFCP2) as critical regulators of CPEB1 expression. Hypermethylation of the CPEB1 promoter resulted in a simultaneous increase in the capacity for TFCP2 binding and a decreased likelihood of CEBPB binding, both of which led to diminished expression of CPEB1. CONCLUSIONS: Our results identified a novel tumour-suppressive role of CPEB1 in CRC and found that hypermethylation of the CPEB1 promoter may lead to diminished expression due to decreased chromatin accessibility and transcription factor binding. Collectively, these results suggest a potential role for CPEB1 in the diagnosis and treatment of CRC.


Asunto(s)
Proteína beta Potenciadora de Unión a CCAAT/genética , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/patología , Metilación de ADN/genética , Proteínas de Unión al ADN/genética , Factores de Transcripción/genética , Factores de Escisión y Poliadenilación de ARNm/genética , Anciano , Línea Celular Tumoral , Epigénesis Genética/genética , Femenino , Regulación Neoplásica de la Expresión Génica/genética , Humanos , Masculino , Persona de Mediana Edad
14.
Mol Ecol Resour ; 21(6): 1983-1995, 2021 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-33709555

RESUMEN

Scale insects are hemimetabolous, showing "incomplete" metamorphosis and no true pupal stage. Ericerus pela, commonly known as the white wax scale insect (hereafter, WWS), is a wax-producing insect found in Asia and Europe. WWS displays dramatic sexual dimorphism, with notably different metamorphic fates in males and females. Males develop into winged adults, while females are neotenic and maintain a nymph-like appearance, which are flightless and remain stationary. Here, we report the de novo assembly of the WWS genome with a size of 638.30 Mbp (69.68 Mbp for scaffold N50) by PacBio sequencing and Hi-C. These data allowed us to perform a robust phylogenetic analysis comprising 24,923 gene orthogroups from 16 representative insect genomes. This analysis indicated that holometabola evolved from insects with incomplete metamorphosis in the Late Carboniferous, about 50 million years earlier than previously thought. To study the distinct developmental fates of males and females, we analysed the methylome landscape in either sex. Surprisingly, WWS displayed high methylation levels (4.42% for males) when compared to other insects. We observed differential methylation patterns in males and females for genes involved in steroid and sesquiterpenoid production as well as genes acting in fatty acid metabolism pathways. We measured titre profiles for ecdysone, the principal insect steroid hormone, and juvenile hormone (a sesquiterpenoid) in both males and females, which suggested that these hormones are the primary drivers of sexually dimorphic development. Our results provide a comprehensive genomic and epigenomic resource of scale insects that provide new insights into the evolution of metamorphosis and sexual dimorphism in insects.


Asunto(s)
Epigenoma , Genoma de los Insectos , Hemípteros/fisiología , Diferenciación Sexual , Animales , Femenino , Hemípteros/genética , Masculino , Metamorfosis Biológica/genética , Filogenia
15.
Sci Rep ; 11(1): 1562, 2021 01 15.
Artículo en Inglés | MEDLINE | ID: mdl-33452296

RESUMEN

The North Pacific subtropical gyre (NPSG) plays a major role in present global ocean circulation. At times, the gyre has coursed through the South China Sea, but its role in the evolutionary development of that Sea remains uncertain. This work systematically describes a major shift in NPSG paleo-circulation evident from sedimentary features observed in seismic and bathymetric data. These data outline two contourite depositional systems-a buried one formed in the late Miocene, and a latest Miocene to present-day system. The two are divided by a prominent regional discontinuity that represents a major shift in paleo-circulation during the latest Miocene (~ 6.5 Ma). The shift coincides with the further restriction of the South China Sea with respect to the North Pacific due to the formation of the Luzon Strait as a consequence of further northwest movement of the Philippine Sea plate. Before that restriction, data indicate vigorous NPSG circulation in the South China Sea. Semi-closure, however, established a new oceanographic circulation regime in the latest Miocene. This work demonstrates the significant role of recent plate tectonics, gateway development, and marginal seas in the establishment of modern global ocean circulation.

16.
Biomed Pharmacother ; 136: 111241, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-33485069

RESUMEN

Previously, we have demonstrated that policosanol from Chinese wax suppressed testosterone(T)-induced alopecia in mice. However, the underlying mechanism remained to be determined. Herein, we investigated the mechanism of policosanol against androgenetic alopecia (AGA). AGA was induced in Kunming mice by subcutaneous administration of testosterone propionate for 60 d. Policosanol (0.5 %, 1% or 2%) was applied topically on the back of mice. Finasteride (2%) was applied topically as a positive control. The serum T and estradiol (E2) concentrations were determined by ELISA after 28 and 60 days of treatment. The cutaneous expression or activity of key mediators of hair growth, such as alkaline phosphatase (ALP), vascular endothelial growth factor (VEGF), and epidermal growth factor (EGF), was measured. MTS assay was performed to evaluate cell proliferation in cultured human dermal papilla cells (DPCs) treated with dihydrotestosterone (DHT). Western blotting was performed to evaluate the protein expression of Bax, Bcl2, TGF-ß2, caspase-9, and caspase-3. We found lower T and T/E2 ratio in mice treated with policosanol than in the model group. Policosanol suppressed premature hair follicle entry into the regression phase, as shown by improving VEGF and EGF expression and ALP activity. The MTS assay showed that policosanol markedly inhibited the apoptosis of DHT-treated DPCs. Western blotting showed that policosanol significantly reduced the protein expression of TGF-ß2, cleaved caspese-9, cleaved caspase-3, and Bax, and increased that of Bcl2. The optimal effect was obtained with 12.50 g/mL policosanol. In conclusion, policosanol prevents androgenetic alopecia by regulating hormone levels and suppressing premature hair follicle entry into the regression phase.


Asunto(s)
Alopecia/tratamiento farmacológico , Alcoholes Grasos/farmacología , Folículo Piloso/efectos de los fármacos , Hemípteros , Fosfatasa Alcalina/metabolismo , Alopecia/sangre , Alopecia/inducido químicamente , Alopecia/fisiopatología , Animales , Apoptosis/efectos de los fármacos , Proteínas Reguladoras de la Apoptosis/metabolismo , Proliferación Celular/efectos de los fármacos , Citocinas/metabolismo , Modelos Animales de Enfermedad , Factor de Crecimiento Epidérmico/metabolismo , Estradiol/sangre , Alcoholes Grasos/aislamiento & purificación , Folículo Piloso/crecimiento & desarrollo , Folículo Piloso/metabolismo , Hemípteros/química , Masculino , Ratones , Testosterona/sangre , Propionato de Testosterona , Factor de Crecimiento Transformador beta2/metabolismo , Factor A de Crecimiento Endotelial Vascular/metabolismo , Ceras
17.
Onco Targets Ther ; 13: 8441-8450, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32922034

RESUMEN

BACKGROUND: Small nucleolar RNA host gene (SNHG) family members are newly recognized lncRNAs, which have been revealed to be oncogenes in several cancers. However, little studies investigated the expression and clinical implications of SNHGs in AML. METHODS: Herein, we systemically determined the prognostic role of the expression of SNHG family members in acute myeloid leukemia (AML). RESULTS: Among the expression of all SNHG family members, we identified that only SNHG7 and SNHG12 expression were found to have prognostic effects on overall survival (OS) and leukemia-free survival (LFS) in AML by Cox regression univariate analysis. Furthermore, Kaplan-Meier analysis showed that SNHG7 higher-expressed cases had markedly longer OS and LFS time than SNHG7 lower-expressed cases, whereas SNHG12 higher-expressed cases had markedly shorter OS and LFS time than SNHG12 lower-expressed cases. Interestingly, SNHG7 and SNHG12 expression were also associated with several prognosis-related clinical/molecular features such as white blood cell counts, FAB/cytogenetic classifications, IDH1 mutation, RUNX1 mutation, and NPM1 mutation. Despite the associations, Cox regression multivariate analysis confirmed the independent prognostic impact of SNHG7 and SNHG12 expression in AML. Notably, we further validated that both SNHG7 and SNHG12 expression was significantly increased in newly diagnosed AML patients. CONCLUSION: Our findings demonstrated that SNHG7 and SNHG12 expression act as independent prognostic indicators in AML.

18.
In Vitro Cell Dev Biol Anim ; 56(6): 444-451, 2020 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-32737835

RESUMEN

In this paper, four established cell lines derived from newly hatched larvae of Papilio demoleus Linnaeus and 57 single-cell clones derived from the 3 lines were used as test materials. Recombinant ß-galactosidase baculovirus AcMNPV-Gal was used to infect the P. demoleus L. cell lines and the single-cell clones, and recombinant protein expression in each cell line was detected and compared. Three clonal cell lines, RIRI-PaDe-1-C1, RIRI-PaDe-2-C6 and RIRI-PaDe-3-C52, which showed significantly higher ß-galactosidase expression levels than those of the parental cell lines, were selected. Five types of commercial serum-free media for insect cells, Express Five SFM, Ex-Cell 405, Sf-900III SFM, Sf-900II SFM and HyClone Serum-Free Media, were used to adapt RIRI-PaDe-2-C6 cells and RIRI-PaDe-3-C52 cells to serum-free culture conditions, and the growth characteristics of the cells and the exogenous protein expression characteristics before and after adaptation were compared. The results showed that RIRI-PaDe-2-C6 cells could stably proliferate in Ex-Cell 405, RIRI-PaDe-3-C52 cells could stably proliferate in Express Five SFM and Ex-Cell 405, and the rate of proliferation of and the level of expression of ß-galactosidase in RIRI-PaDe-3-C52 cells were significantly increased in Express Five SFM.


Asunto(s)
Adaptación Fisiológica , Mariposas Diurnas/citología , Células Clonales/citología , Expresión Génica , Análisis de la Célula Individual , beta-Galactosidasa/metabolismo , Animales , Baculoviridae , Línea Celular , Proliferación Celular , Forma de la Célula , Medio de Cultivo Libre de Suero , Cinética , Proteínas Recombinantes/metabolismo
19.
Front Genet ; 11: 511, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32528529

RESUMEN

Major histocompatibility complex (HLA) class I chain-related protein A (MICA) regulates immune surveillance through activation of NKG2D (natural killer group 2D) receptor. However, the genetic association, potential function, and predictive ability of MICA alleles with colorectal cancer (CRC) prognosis remain undefined. In this study, we characterized MICA alleles in tissue samples from 104 patients with CRC and 536 healthy controls and carried out genetic association studies by molecular and clinical CRC phenotypes. Preliminary sequence analysis revealed that MICA ∗009:01 or ∗049 alleles were significantly decreased in patients with CRC (p = 0.0049), and further stratification analysis indicated that MICA ∗012:01 allele was associated with patients with CRC and carrying KRAS codon 12 mutation (p = 0.027). The functional consequences of MICA alleles were examined via transfected CRC cell lines which showed that overexpression of MICA ∗012:01 enhanced the proliferation, invasion, and metastatic phenotype of CRC. Preliminary analysis of disease-free survival time in patients with and without MICA ∗012:01 suggest this allele may be predictive for poor prognosis of patients with KRAS codon 12 mutated CRC, as no somatic mutation of MICA gene was detected in CRC tumors compared to paracancerous tissues. Our study indicates that MICA ∗012:01 allele is associated with KRAS-mutated CRC, facilitates CRC invasion and metastasis, and possibly reduces the survival of patients with KRAS-mutated CRC.

20.
DNA Cell Biol ; 38(10): 1048-1055, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31433200

RESUMEN

DNA condensed agents can improve the transfection efficiency of the cationic liposome delivery system. However, various condensed agents have distinct transfection efficiency and cellular cytotoxicity. The object of this study was to screen the optimal agents with the high transfection efficiency and low cytotoxicity from four polymer compressive materials, polyethylenimine (PEI), chitosan, poly-l-lysine (PLL), and spermidine. DNA was precompressed with these four agents and then combined to cationic liposomes. Subsequently, the entrapment and transfection efficiency of the obtained complexes were investigated. Finally, the particle sizes, cytotoxicity, and endocytosis fashion of these copolymers (Lipo-PEI, Lipo-chitosan, Lipo-PLL, and Lipo-spermidine) were examined. It was found that these four copolymers had significantly lower cytotoxicity and higher transfection efficiency (45.5%, 42.4%, 36.8%, and 47.4%, respectively) than those in the control groups. The transfection efficiency of Lipo-PEI and Lipo-spermidine copolymers were better than the other two copolymers. In 293T cells, nystatin significantly inhibited the transfection efficiency of Lipo-PEI-DNA and Lipo-spermidine-DNA (51.88% and 46.05%, respectively), which suggest that the endocytosis pathway of Lipo-spermidine and Lipo-PEI copolymers was probably caveolin dependent. Our study indicated that these dual-degradable copolymers especially liposome-spermidine copolymer could be used as the potential biocompatible gene delivery carriers.


Asunto(s)
Quitosano/química , Liposomas/química , Polietileneimina/química , Polilisina/química , Espermidina/química , Transfección/métodos , Cationes , Caveolina 1/genética , Caveolina 1/metabolismo , Quitosano/metabolismo , Colesterol/química , Colesterol/metabolismo , Endocitosis/efectos de los fármacos , Endocitosis/genética , Ácidos Grasos Monoinsaturados/química , Ácidos Grasos Monoinsaturados/metabolismo , Células HEK293 , Humanos , Liposomas/metabolismo , Nistatina/farmacología , Tamaño de la Partícula , Plásmidos/química , Plásmidos/metabolismo , Polietileneimina/metabolismo , Polilisina/metabolismo , Compuestos de Amonio Cuaternario/química , Compuestos de Amonio Cuaternario/metabolismo , Espermidina/metabolismo
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