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1.
Front Vet Sci ; 8: 708077, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34805328

RESUMO

Dermatomycosis is the second major cause of morbidity in giant pandas (Ailuropoda melanoleuca), and seriously endangers its health. Previous observations indicated that the occurrence of dermatomycosis in the giant panda varies in different seasons. The skin microbiota is a complex ecosystem, but knowledge on the community structure and the pathogenic potentials of fungi on the skin of the giant panda remains limited. In this study, samples from the giant panda skin in different seasons were collected, and the mycobiota were profiled by 18S rRNA gene sequencing. In total, 375 genera in 38 phyla were detected, with Ascomycota, Basidiomycota, Streptophyta, and Chlorophyta as the predominant phyla and Trichosporon, Guehomyces, Davidiella, Chlorella, Asterotremella, and Klebsormidium as the predominant genera. The skin mycobiota of the giant panda changed in the seasons, and the diversity and abundance of the skin fungi were significantly higher in spring, autumn, and summer than in the winter. Several dermatomycosis-associated fungi were detected as opportunists in the skin mycobiota of healthy giant pandas. Clinical dermatomycosis in the giant panda is observed more in summer and autumn. In this study, the results indicated that the high diversity and abundance of the skin fungi may have enhanced the occurrence of dermatomycosis in autumn and summer, and that dermatomycosis-associated fungi are the normal components of the skin mycobiota.

2.
Int J Mol Sci ; 22(21)2021 Oct 27.
Artigo em Inglês | MEDLINE | ID: mdl-34769012

RESUMO

Ageratina adenophora is one of the major invasive weeds that causes instability of the ecosystem. Research has reported that A. adenophora produces allelochemicals that inhibit the growth and development of food crops, and also contain some toxic compounds that cause toxicity to animals that consume it. Over the past decades, studies on the identification of major toxic compounds of A. adenophora and their toxic molecular mechanisms have been reported. In addition, weed control interventions, such as herbicides application, was employed to reduce the spread of A. adenophora. However, the development of therapeutic and prophylactic measures to treat the various A. adenophora-induced toxicities, such as hepatotoxicity, splenotoxicity and other related disorders, have not been established to date. The main toxic pathogenesis of A. adenophora is oxidative stress and inflammation. However, numerous studies have verified that some extracts and secondary metabolites isolated from A. adenophora possess anti-oxidation and anti-inflammation activities, which implies that these extracts can relieve toxicity and aid in the development of drug or feed supplements to treat poisoning-related disorders caused by A. adenophora. Furthermore, beneficial bacteria isolated from rumen microbes and A. adenophora can degrade major toxic compounds in A. adenophora so as to be developed into microbial feed additives to help ameliorate toxicity mediated by A. adenophora. This review presents an overview of the toxic mechanisms of A. adenophora, provides possible therapeutic strategies that are available to mitigate the toxicity of A. adenophora and introduces relevant information on identifying novel prophylactic and therapeutic measures against A. adenophora-induced toxicity.

3.
J Vis Exp ; (176)2021 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-34779435

RESUMO

Osteosarcoma is the most common primary bone cancer in children and adolescents, with lungs as the most common metastatic site. The five-year survival rate of osteosarcoma patients with pulmonary metastasis is less than 30%.Therefore, the utilization of mouse models mimicking the osteosarcoma development in humans is of great significance for understanding the fundamental mechanism of osteosarcoma carcinogenesis and pulmonary metastasis to develop novel therapeutics. Here, detailed procedures are reported to generate the primary osteosarcoma and pulmonary metastasis mouse models via intratibia injection of osteosarcoma cells. Combined with the bioluminescence or X-ray live imaging system, these living mouse models are utilized to monitor and quantify osteosarcoma growth and metastasis. To establish this model, a basement membrane matrix containing osteosarcoma cells was loaded in a micro-volume syringe and injected into one tibia of each athymic mouse after being anesthetized. The mice were sacrificed when the primary osteosarcoma reached the size limitation in the IACUC-approved protocol. The legs bearing osteosarcoma and the lungs with metastasis lesions were separated. These models are characterized by a short incubation period, rapid growth, severe lesions, and sensitivity in monitoring the development of primary and pulmonary metastatic lesions. Therefore, these are ideal models for exploring the functions and mechanisms of specific factors in osteosarcoma carcinogenesis and pulmonary metastasis, the tumor microenvironment, and evaluating the therapeutic efficacy in vivo.

4.
Comput Intell Neurosci ; 2021: 4529107, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34790231

RESUMO

Frequent occurrence and long-term existence of respiratory diseases such as COVID-19 and influenza require bus drivers to wear masks correctly during driving. To quickly detect whether the mask is worn correctly on resource-constrained devices, a lightweight target detection network SAI-YOLO is proposed. Based on YOLOv4-Tiny, the network incorporates the Inception V3 structure, replaces two CSPBlock modules with the RES-SEBlock modules to reduce the number of parameters and computational difficulty, and adds a convolutional block attention module and a squeeze-and-excitation module to extract key feature information. Moreover, a modified ReLU (M-ReLU) activation function is introduced to replace the original Leaky_ReLU function. The experimental results show that SAI-YOLO reduces the number of network parameters and calculation difficulty and improves the detection speed of the network while maintaining certain recognition accuracy. The mean average precision (mAP) for face-mask-wearing detection reaches 86% and the average precision (AP) for mask-wearing normative detection reaches 88%. In the resource-constrained device Raspberry Pi 4B, the average detection time after acceleration is 197 ms, which meets the actual application requirements.


Assuntos
Condução de Veículo , COVID-19 , Humanos , Reconhecimento Psicológico , SARS-CoV-2
5.
Front Microbiol ; 12: 700704, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34616374

RESUMO

Transportation is an inevitable phase for the cattle industry, and its effect on the respiratory system of transported cattle remains controversial. To reveal cattle's nasopharyngeal microbiota dynamics, we tracked a batch of beef calves purchased from an auction market, transported to a farm by vehicle within 3 days, and adaptively fed for 7 days. Before and after the transport and after the placement, a total of 18 nasopharyngeal mucosal samples were collected, and microbial profiles were obtained using a metagenomic shotgun sequencing approach. The diversity, composition, structure, and function of the microbiota were collected at each time point, and their difference was analyzed. The results showed that, before the transportation, there were a great abundance of potential bovine respiratory disease (BRD)-related pathogens, and the transportation did not significantly change their abundance. After the transportation, 7 days of placement significantly decreased the risk of BRD by decreasing the abundance of potential BRD-related pathogens even if the diversity was decreased. We also discussed the controversial results of transportation's effect in previous works and the decrease in diversity induced by placement. Our work provided more accurate information about the effect of transportation and the followed placement on the calf nasopharyngeal microbial community, indicated the importance of adaptive placement after long-distance transport, and is helpful to prevent BRD induced by transportation stress.

6.
Funct Integr Genomics ; 21(5-6): 695-707, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34676472

RESUMO

Haemophilus (Glaesserella) parasuis is a commensal bacterium that causes Glässer's disease (GD) in swine. As a global transcriptional factor, CheY regulates the expression of hundreds of genes in H. parasuis. In this study, we measured changes in gene expression at the whole transcriptome level using RNAseq. We identified 2058 co-expressed genes, and found 624 differentially expressed genes (q < 0.05) in ΔcheY and SC1401. Several important GO annotations and signaling pathways were identified. RNA-seq results were assembled according to the reference genome, compared with the annotated gene model, and 12 new transcriptional regions were found. Finally, q-PCR results validated the RNA-seq results with 8 randomly selected genes. The present study indicated that CheY is mainly involved in the regulation of ABC transport, oxidative phosphorylation, and ß-Lactam resistance. We draw the regulatory network of CheY, which offers greater insight into the regulatory mechanism of CheY in H.parasuis.

7.
Int J Mol Sci ; 22(18)2021 Sep 17.
Artigo em Inglês | MEDLINE | ID: mdl-34576206

RESUMO

Actinobacillus pleuropneumoniae is a pathogen that infects pigs and poses a serious threat to the pig industry. The emergence of quinolone-resistant strains of A.pleuropneumoniae further limits the choice of treatment. However, the mechanisms behind quinolone resistance in A.pleuropneumoniae remain unclear. The genomes of a ciprofloxacin-resistant strain, A. pleuropneumoniae SC1810 and its isogenic drug-sensitive counterpart were sequenced and analyzed using various bioinformatics tools, revealing 559 differentially expressed genes. The biological membrane, plasmid-mediated quinolone resistance genes and quinolone resistance-determining region were detected. Upregulated expression of efflux pump genes led to ciprofloxacin resistance. The expression of two porins, OmpP2B and LamB, was significantly downregulated in the mutant. Three nonsynonymous mutations in the mutant strain disrupted the water-metal ion bridge, subsequently reducing the affinity of the quinolone-enzyme complex for metal ions and leading to cross-resistance to multiple quinolones. The mechanism of quinolone resistance in A. pleuropneumoniae may involve inhibition of expression of the outer membrane protein genes ompP2B and lamB to decrease drug influx, overexpression of AcrB in the efflux pump to enhance its drug-pumping ability, and mutation in the quinolone resistance-determining region to weaken the binding of the remaining drugs. These findings will provide new potential targets for treatment.


Assuntos
Quinolonas/farmacologia , Actinobacillus pleuropneumoniae/efeitos dos fármacos , Actinobacillus pleuropneumoniae/genética , Biofilmes/efeitos dos fármacos , Porinas/metabolismo , Transcriptoma/genética
8.
In Vitro Cell Dev Biol Anim ; 57(7): 685-694, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34518994

RESUMO

The destruction of biological activity such as senescence and apoptosis caused by oxidative stress could play a pivotal role in the poor therapeutic efficiency of bone marrow mesenchymal stem cells (BMSCs) transplantation. Mitoquinone (MitoQ) has a highly effective mitochondrial antioxidant effect, and has been widely used in many oxidative damage models. This study aimed to investigate the protective effect of MitoQ on the oxidative stress-mediated senescence of canine BMSCs and the underlying mechanism. The senescence of BMSCs was determined by senescence-associated ß-galactosidase staining and quantitative real-time PCR. The expression of p-Nrf2 protein was detected by Western blotting. The results demonstrated that, as BMSCs were expanded in vitro, the senescent phenotype appeared. And the senescence of BMSCs may be caused by oxidative stress, manifested by increasing the level of ROS and decreasing the activity of antioxidant enzymes. Treatment of MitoQ down-regulated the mRNA levels of senescence-related and apoptosis-related genes, but up-regulated the mRNA levels of proliferation-related genes. Meanwhile, ROS generation and senescent activity were reduced in MitoQ-treated BMSCs. Further mechanism studies showed that MitoQ obviously promoted Nrf2 phosphorylation, and also facilitated the translocation of Nrf2 into the nucleus. Moreover, treatment of MitoQ increased the mRNA levels of downstream antioxidant genes and enhanced the activities of superoxide dismutase, catalase, and glutathione peroxidase. Thus, our study revealed that MitoQ, via the Nrf2/ARE signaling pathway, exerts an antioxidant effect as well as potentially delays OS-mediated senescence during BMSCs that were expanded in vitro, which may serve as a novel strategy to optimize the clinical application of BMSCs.

9.
Front Microbiol ; 12: 707548, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34557168

RESUMO

Bacterial infection and imbalance of bacterial community in the genitourinary system of giant panda could affect the reproductive health. In severe cases, it can also lead to abortion. In this study, 13 of vaginal secretions in the estrue (E) group and seven of vaginal secretions in the non-estrue (NE) group were used to study the composition and diversity of vaginal bacterial communities between estrus and non-estrus by 16S rRNA gene sequencing analysis. The results showed that the vaginal microbiome in giant pandas shared the same top five abundant species between estrus and non-estrus at the phylum level. However, the vaginal microbiome changed significantly during estrus at the genus level. In top 10 genera, the abundance of Escherichia, Streptococcus, and Bacteroides in the E group was significantly higher than that in the NE group (p<0.05); Azomonas, Porphyromonas, Prevotella, Campylobacter, and Peptoniphilus in the NE group was significantly higher than that in the E group (p<0.05). The richness and diversity of vaginal microbiome in giant panda on estrus were significantly lower than those on non-estrus (p<0.05). It is noteworthy that the abundance of Streptococcus, Escherichia, and Bacteroides of vagina in giant pandas maintained low abundance in the daily. Whereas, they increased significantly during estrus period, which may play an important role in female giant pandas during estrus period. It was hypothesized that hormones may be responsible for the changes in the vaginal microbiome of giant pandas between estrus and no-estrus stages.

10.
Sci Total Environ ; 798: 149268, 2021 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-34333432

RESUMO

Antimicrobial resistance (AMR) has become a public health concern; but antibiotic resistance genes (ARGs) and integrons that link to AMR of Escherichia coli from non-human primates remain largely unknown. This study aimed to investigate antibiotic resistance, emerging environmental pollutants ARGs, and integrons factors (intI1, intI2 and intI3) in 995 E. coli isolates obtained from 50 species of captive non-human primates of 13 zoos in China. Our result showed 83.62% of the E. coli isolates were resistant to at least one antibiotic and 47.94% isolates showed multiple drug resistances (MDR). The E. coli isolates mainly showed resistance to tetracycline (tetracycline 62.71%, doxycycline 61.11%), ß-lactams (ampicillin 54.27%, amoxicillin 52.36%), and sulfonamide (trimethoprim-sulfamethoxazole 36.78%). A total of 423 antibiotic resistance patterns were observed, of which DOX/TET (49 isolates, 4.92%) was the most common pattern. Antibiotic resistance rates among 13 zoos had a significant difference (P < 0.01). We further detected 22 ARGs in the 995 E. coli isolates, of which tetA had the highest occurrence (70.55%). The presence of integrons class 1 and 2 were 24.22% and 1.71%, respectively, while no class 3 integron was found. Significant positive associations were observed among integrons and antibiotics, of which the strongest association was observed for integrons / Gentamicin (OR, 2.642) and integrons / Cefotaxime (OR, 2.512). In addition, cassette arrays were detected in 64 strains of class 1 integron-positive isolates (26.56%) and 10 strains of class 2 integron-positive isolates (58.82%). Eighteen cassette arrays were found within 64 class 1 integron isolates, while 3 cassette arrays were identified within 10 class 2 integron isolates. Our results indicate a high diversity of antibiotic resistance phenotypes in non-human primate E. coli isolates, which carry multiple ARGs and integrons. Corresponding preventive measures should be taken to prevent the spread of integron-mediated ARGs in non-human primates and their living environments in zoos.


Assuntos
Infecções por Escherichia coli , Integrons , Animais , Antibacterianos/farmacologia , Farmacorresistência Bacteriana/genética , Farmacorresistência Bacteriana Múltipla/genética , Escherichia coli/genética , Infecções por Escherichia coli/epidemiologia , Integrons/genética , Testes de Sensibilidade Microbiana , Prevalência , Primatas
11.
Front Vet Sci ; 8: 666486, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34291099

RESUMO

The giant panda is one of the rarest animals in the world. Skin diseases seriously endanger the health of giant panda and are considered the second major cause of its morbidity. Skin microbiota is a complex ecosystem, and the community structure and the pathogenic potential of bacteria on giant panda skin remain largely unclear. In order to understand the skin bacterial flora of captive giant pandas, the microbiota in giant panda skin samples collected during different seasons was profiled via 16S rRNA gene sequencing. In total, 522 genera from 53 bacterial phyla were detected, with Proteobacteria (40.5%), Actinobacteria (23.1%), Firmicutes (21.1%), Bacteroidetes (9.5%), Cyanobacteria (2.1%), and Thermi (1.2%) as the predominant phyla and Streptococcus (13.9%), Acinetobacter (9.2%), Staphylococcus (2.9%), Pseudomonas (5.9%), Dermacoccus (4.8%), Brachybacterium (2.9%), Escherichia (2.7%), Chryseobacterium (2.1%), Arthrobacter (1.6%), Kocuria (1.5%), Psychrobacter (1.2%), Deinococcus (1.1%), and Flavobacterium (1.1%) as the predominant genera. The results indicated that the diversity was lower in winter than in other seasons and higher in autumn than in other seasons, and the abundance in spring was significantly higher than that in other seasons. Several skin disease-associated bacteria were detected as opportunists in the skin microbiota of healthy giant pandas. In this study, the results indicated that the high diversity and abundance of the skin bacteria may have enhanced the occurrence of skin disease in autumn and spring and that skin disease-associated bacteria are the normal components of the skin microbiota.

12.
Int J Biol Sci ; 17(10): 2504-2522, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34326690

RESUMO

Background: As the leading primary bone cancer in adolescents and children, osteosarcoma patients with metastasis show a five-year-survival-rate of 20-30%, without improvement over the past 30 years. Wnt/ß-catenin is important in promoting osteosarcoma development. DKK3 is a Wnt/ß-catenin antagonist and predicted to have the specific binding site in 3'-UTR with miR-214-3p. Methods: miR-214-3p and DKK3 levels were investigated in human osteosarcoma tissues and cells by RT-qPCR; the prognostic importance of DKK3 level in osteosarcoma patients was determined with Log-rank test; direct binding between DKK3 with miR-214-3p was identified with targetscan; anti-osteosarcoma mechanism of cantharidin was investigated by miR-214-3p silence/over-expression with or without cantharidin treatment, and nuclear/cytoplasmic protein assay in osteosarcoma cells. Results: Down-regulated DKK3 indicated poor prognosis of osteosarcoma patients. Up-regulated miR-214-3p promoted proliferation and migration, while suppressed apoptosis of osteosarcoma cells by increasing ß-catenin nuclear translocation and LEF1 translation via degradation of DKK3. Cantharidin suppressed viabilities, migration and invasion, while promoted cell cycle arrest and apoptosis in 143B and U-2 OS cells via down-regulating miR-214-3p to up-regulate DKK3, thus inhibited p-GSK-3ß expression, ß-catenin nuclear translocation and LEF1 translation. Meanwhile, cantharidin inhibited tumor growth in xenograft-bearing mice with 143B cell injection in tibia. Conclusion: miR-214-3p mediated Wnt/ß-catenin/LEF1 signaling activation by targeting DKK3 to promote oncogenesis of osteosarcoma; cantharidin inhibited proliferation and metastasis of osteosarcoma cells via down-regulating miR-214-3p to up-regulate DKK3 and decrease ß-catenin nuclear translocation, indicating that cantharidin may be a prospective candidate for osteosarcoma treatment by targeting miR-214-3p/DKK3/ß-catenin signaling.

13.
Molecules ; 26(11)2021 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-34204150

RESUMO

The purpose of this study was to develop mixed polymeric micelles with high drug loading capacity to improve the oral bioavailability of icaritin with Soluplus® and Poloxamer 407 using a creative acid-base shift (ABS) method, which exhibits the advantages of exclusion of organic solvents, high drug loading and ease of scaling-up. The feasibility of the ABS method was successfully demonstrated by studies of icaritin-loaded polymeric micelles (IPMs). The prepared IPMs were characterized to have a spherical shape with a size of 72.74 ± 0.51 nm, and 13.18% drug loading content. In vitro release tests confirmed the faster release of icaritin from IPMs compared to an oil suspension. Furthermore, bioavailability of icaritin in IPMs in beagle dogs displayed a 14.9-fold increase when compared with the oil suspension. Transcellular transport studies of IPMs across Caco-2 cell monolayers confirmed that the IPMs were endocytosed in their intact forms through macropinocytosis, clathrin-, and caveolae-mediated pathways. In conclusion, the results suggested that the mixed micelles of Soluplus® and Poloxamer 407 could be a feasible drug delivery system to enhance oral bioavailability of icaritin, and the ABS method might be a promising technology for the preparation of polymeric micelles to encapsulate poorly water-soluble weakly acidic and alkaline drugs.


Assuntos
Flavonoides/administração & dosagem , Poloxâmero/química , Polietilenoglicóis/química , Polivinil/química , Transdução de Sinais/efeitos dos fármacos , Administração Oral , Animais , Disponibilidade Biológica , Células CACO-2 , Cavéolas/metabolismo , Clatrina/metabolismo , Cães , Estudos de Viabilidade , Flavonoides/síntese química , Flavonoides/farmacocinética , Humanos , Masculino , Micelas , Nanopartículas , Tamanho da Partícula
15.
Infect Genet Evol ; 92: 104912, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-33989813

RESUMO

Giardia duodenalis is a common protozoan parasite that can infect humans and animals. Although previous studies demonstrated that the assemblage E of G. duodenalis is prevalent in cattle, studies on its genetic diversity were mostly based on single loci and very few involved multilocus analysis. To better understand the genetic variability and structure of G. duodenalis assemblage E in Chinese dairy cattle, 651 multilocus sequences derived from nine provinces (Gansu, Guangdong, Henan, Jiangsu, Ningxia, Shaanxi, Shanghai, Sichuan and Xinjiang) of China were analyzed in this study. Results showed that a total of 220 haplotypes were identified in the G. duodenalis assemblage E, with a high haplotype diversity (Hd = 0.97225) and low nucleotide diversity (π = 0.00259). The genetic differentiation index (FST) and gene flow (Nm) results indicated low degree of genetic differentiation, implying frequent genetic communication. Combined with the analysis of molecular variance (AMOVA), genetic variation within populations (81.7%) was higher than that among populations (18.3%), indicating low degree of genetic differentiation between populations. Such low rates of gene differentiation supported no significant correlations with geographical divisions. Moreover, both negative Tajima's D and Fu's FS values of neutrality tests and unimodal curve of mismatch distribution analyses indicated that G. duodenalis assemblage E population in Chinese dairy cattle had experienced demographic expansion. Overall, these findings contribute to an improved understanding of the population genetics and evolutionary biology of G. duodenalis assemblage E and assist in its control in cattle.

16.
Parasite ; 28: 31, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33812463

RESUMO

Enterocytozoon bieneusi is a common intracellular parasite that infects a wide range of hosts, including humans and companion animals, raising concerns of zoonotic transmission. However, there is limited epidemiological information on the prevalence and genotypes of E. bieneusi in sheltered dogs and cats in Sichuan province, southwestern China. A total of 880 fecal samples were collected from shelters in different cities of Sichuan province, including 724 samples from dogs, and 156 samples from cats. Enterocytozoon bieneusi was determined by sequence analysis of the ribosomal internal transcribed spacer (ITS). Overall, the prevalence of E. bieneusi was 18% (158/880), and the parasite was detected in 18.8% (136/724) and 14.1% (22/156) of the dogs and cats examined, respectively. Sequence analysis revealed the presence of five genotypes in dogs, including three known genotypes CD9 (n = 92), PtEb IX (n = 41), and Type IV (n = 1), and two novel genotypes SCD-1 (n = 1) and SCD-2 (n = 1). Similarly, four genotypes were identified in cats, including CD9 (n = 11), Type IV (n = 6), D (n = 4), and PtEb IX (n = 1). Genotypes D and Type IV have previously been identified in humans and are reported in sheltered dogs and cats in the present study, indicating that these animals could be as potential sources of human microsporidiosis infections.


Assuntos
Doenças do Gato , Doenças do Cão , Enterocytozoon , Microsporidiose , Animais , Doenças do Gato/epidemiologia , Gatos , China/epidemiologia , Doenças do Cão/epidemiologia , Cães , Enterocytozoon/genética , Fezes , Genótipo , Microsporidiose/epidemiologia , Microsporidiose/veterinária , Filogenia , Prevalência , Zoonoses
17.
Eur J Med Chem ; 217: 113381, 2021 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-33756124

RESUMO

KRAS plays an essential role in regulating cell proliferation, differentiation, migration and survival. Mutated KRAS is a major driver of malignant transformation in multiple human cancers. We showed previously that fendiline (6) is an effective inhibitor of KRAS plasma membrane (PM) localization and function. In this study, we designed, synthesized and evaluated a series of new fendiline analogs to optimize its drug properties. Systemic structure-activity relationship studies by scaffold repurposing led to the discovery of several more active KRAS PM localization inhibitors such as compounds 12f (NY0244), 12h (NY0331) and 22 (NY0335) which exhibit nanomolar potencies. These compounds inhibited oncogenic KRAS-driven cancer cell proliferation at single-digit micromolar concentrations in vitro. In vivo studies in a xenograft model of pancreatic cancer revealed that 12h and 22 suppressed oncogenic KRAS-expressing MiaPaCa-2 tumor growth at a low dose range of 1-5 mg/kg with no vasodilatory effects, indicating their potential as chemical probes and anticancer therapeutics.


Assuntos
Antineoplásicos/farmacologia , Membrana Celular/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Fendilina/farmacologia , Proteínas Proto-Oncogênicas p21(ras)/antagonistas & inibidores , Animais , Antineoplásicos/síntese química , Antineoplásicos/química , Membrana Celular/metabolismo , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Cães , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/química , Feminino , Fendilina/análogos & derivados , Fendilina/química , Humanos , Camundongos , Camundongos Nus , Estrutura Molecular , Neoplasias Experimentais/tratamento farmacológico , Neoplasias Experimentais/metabolismo , Neoplasias Experimentais/patologia , Proteínas Proto-Oncogênicas p21(ras)/metabolismo , Relação Estrutura-Atividade
18.
Exp Ther Med ; 21(5): 436, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33777189

RESUMO

Cataracts account for ~50% of the cases of blindness in individuals worldwide. The apoptosis of lens epithelial cells (LECs) occurs during the formation of cataracts, which is a non-congenital condition. Numerous microRNAs (miRs) have been reported to regulate apoptosis in LECs. For instance, miR-23a expression levels were shown to be upregulated in cataractous lenses; however, the function of miR-23a in cataracts remains undetermined. To establish an in vitro model of cataracts, human LECs, HLE-B3 cells, were induced with 200 µmol/l H2O2 for 24 h. HLE-B3 cells were transfected with the miR-negative control (NC) mimic, miR-23a-3p mimic, miR-NC inhibitor, miR-23a-3p inhibitor, small interfering RNA (siRNA) targeting BCL2 (siRNA-BCL2) and siRNA-NC. The expression levels of miR-23a-3p were detected using reverse transcription-quantitative PCR. The interaction between miR-23a-3p and the 3'-untranslated region (UTR) of the target mRNA BCL2 was predicted by TargetScan 7.1, and further validated using a dual luciferase reporter assay. The BCL2 protein expression levels were analyzed using western blotting, cell proliferation was determined using a CCK-8 assay and the levels of cell apoptosis were analyzed using flow cytometric analysis. The results of the present study revealed that the expression levels of miR-23a-3p were significantly upregulated, while the expression levels of BCL2 were significantly downregulated in H2O2-induced HLE-B3 cells compared to untreated control cells. BCL2 was shown to be a target of miR-23a-3p. The miR-23a-3p inhibitor subsequently attenuated H2O2-induced apoptosis and increased the proliferation of HLE-B3 cells, which was partially reversed by siRNA-BCL2. In conclusion, the findings of the current study suggested that the inhibition of miR-23a-3p may attenuate H2O2-induced cataract formation by targeting BCL2, thus providing a novel therapeutic target for the treatment of patients with cataracts in the clinic.

19.
Virulence ; 12(1): 520-546, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-33525975

RESUMO

Polyamines are small, polycationic molecules with a hydrocarbon backbone and multiple amino groups required for optimal cell growth. The potD gene, belonging to the ABC (ATP-binding cassette) transport system potABCD, encodes the bacterial substrate-binding subunit of the polyamine transport system, playing a pivotal role in bacterial metabolism and growth. The swine pathogen Glaesserella parasuis possesses an intact pot operon, and the studies presented here mainly examined the involvement of PotD in Glaesserella pathogenesis. A potD-deficient mutant was constructed using a virulent G. parasuis strain SC1401 by natural transformation; immuno-electron microscopy was used to identify the subcellular location of native PotD protein; an electron microscope was adopted to inspect biofilm and bacterial morphology; immunofluorescence technique was employed to study cellular adhesion, the levels of inflammation and apoptosis. The TSA++-pre-cultured mutant strain showed a significantly reduced adhesion capacity to PK-15 and MLE-12 cells. Likewise, we also found attenuation in virulence using murine models focusing on the clinical sign, H&E, and IFA for inflammation and apoptosis. However, when the mutant was grown in TSB++, virulence recovered to normal levels, along with a high level of radical oxygen species formation in the host. The expression of PotD could actively stimulate the production of ROS in Raw 264.7. Our data suggested that PotD from G. parasuis has a high binding potential to polyamine, and is essential for the full bacterial virulence within mouse models. However, the virulence of the potD mutant is highly dependent on its TSA++ culture conditions rather than on biofilm-formation.


Assuntos
Proteínas de Bactérias/genética , Biofilmes/crescimento & desenvolvimento , Haemophilus parasuis/genética , Haemophilus parasuis/patogenicidade , Proteínas de Membrana Transportadoras/genética , Poliaminas/metabolismo , Animais , Proteínas de Bactérias/metabolismo , Modelos Animais de Doenças , Feminino , Deleção de Genes , Infecções por Haemophilus/microbiologia , Proteínas de Membrana Transportadoras/metabolismo , Camundongos , Organismos Livres de Patógenos Específicos , Virulência/genética
20.
Sci Rep ; 11(1): 2699, 2021 01 29.
Artigo em Inglês | MEDLINE | ID: mdl-33514801

RESUMO

Expanded newborn screening facilitates early identification and intervention of patients with inborn errors of metabolism (IEMs), There is a lack of disease spectrum data for many areas in China. To determine the disease spectrum and genetic characteristics of IEMs in Xi'an city of Shaanxi province in northwest China, 146152 newborns were screening by MSMS from January 2014 to December 2019 and 61 patients were referred to genetic analysis by next generation sequencing (NGS) and validated by Sanger sequencing. Seventy-five newborns and two mothers were diagnosed with IEMs, with an overall incidence of 1:1898 (1:1949 without mothers). There were 35 newborns with amino acidemias (45.45%, 1:4176), 28 newborns with organic acidurias (36.36%, 1:5220), and 12 newborns and two mothers with FAO disorders (18.18%; 1:10439 or 1:12179 without mothers). Phenylketonuria and methylmalonic acidemia were the two most common disorders, accounting for 65.33% (49/75) of all confirmed newborn. Some hotspot mutations were observed for several IEMs, including PAH gene c.728G>A for phenylketonuria; MMACHC gene c.609G>A and c.567dupT, MMUT gene c.323G>A for methylmalonic acidemia and SLC25A13 gene c.852_855del for citrin deficiency. Our study provides effective clinical guidance for the popularization and application of expanded newborn screening, genetic screening, and genetic counseling of IEMs in this region.


Assuntos
Erros Inatos do Metabolismo dos Aminoácidos , Sequenciamento de Nucleotídeos em Larga Escala , Mutação , Triagem Neonatal , Fenilcetonúrias , Erros Inatos do Metabolismo dos Aminoácidos/epidemiologia , Erros Inatos do Metabolismo dos Aminoácidos/genética , China/epidemiologia , Feminino , Humanos , Incidência , Recém-Nascido , Masculino , Fenilcetonúrias/epidemiologia , Fenilcetonúrias/genética
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