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1.
Brief Bioinform ; 25(3)2024 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-38627939

RESUMO

The latest breakthroughs in spatially resolved transcriptomics technology offer comprehensive opportunities to delve into gene expression patterns within the tissue microenvironment. However, the precise identification of spatial domains within tissues remains challenging. In this study, we introduce AttentionVGAE (AVGN), which integrates slice images, spatial information and raw gene expression while calibrating low-quality gene expression. By combining the variational graph autoencoder with multi-head attention blocks (MHA blocks), AVGN captures spatial relationships in tissue gene expression, adaptively focusing on key features and alleviating the need for prior knowledge of cluster numbers, thereby achieving superior clustering performance. Particularly, AVGN attempts to balance the model's attention focus on local and global structures by utilizing MHA blocks, an aspect that current graph neural networks have not extensively addressed. Benchmark testing demonstrates its significant efficacy in elucidating tissue anatomy and interpreting tumor heterogeneity, indicating its potential in advancing spatial transcriptomics research and understanding complex biological phenomena.


Assuntos
Benchmarking , Perfilação da Expressão Gênica , Análise por Conglomerados , Redes Neurais de Computação
2.
J Environ Manage ; 359: 120954, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38692026

RESUMO

Plastic products' widespread applications and their non-biodegradable nature have resulted in the continuous accumulation of microplastic waste, emerging as a significant component of ecological environmental issues. In the field of microplastic detection, the intricate morphology poses challenges in achieving rapid visual characterization of microplastics. In this study, photoacoustic imaging technology is initially employed to capture high-resolution images of diverse microplastic samples. To address the limited dataset issue, an automated data processing pipeline is designed to obtain sample masks while effectively expanding the dataset size. Additionally, we propose Vqdp2, a generative deep learning model with multiple proxy tasks, for predicting six forms of microplastics data. By simultaneously constraining model parameters through two training modes, outstanding morphological category representations are achieved. The results demonstrate Vqdp2's excellent performance in classification accuracy and feature extraction by leveraging the advantages of multi-task training. This research is expected to be attractive for the detection classification and visual characterization of microplastics.


Assuntos
Aprendizado Profundo , Microplásticos , Técnicas Fotoacústicas , Microplásticos/análise , Técnicas Fotoacústicas/métodos , Monitoramento Ambiental/métodos , Plásticos
3.
BMC Cancer ; 23(1): 1114, 2023 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-37968576

RESUMO

BACKGROUND: The establishment of sister chromatid cohesion N-acetyltransferase 2 (ESCO2) is involved in the development of multiple malignancies. However, its role in hypopharyngeal carcinoma (HPC) progression remains uncharacterized. METHODS: This study employed bioinformatics to determine the ESCO2 expression in head and neck squamous cell carcinoma (HNSC) and normal tissues. In vitro cell proliferation, migration, apoptosis, and/or cell cycle distribution assays were used to determine the function of ESCO2 and its relationship with STAT1. Xenograft models were established in nude mice to determine ESCO2 in HPC growth in vivo. Co-immunoprecipitation/mass spectrometry (Co-IP/MS) was conducted to identify the potential ESCO2 binding partners. RESULTS: We found that ESCO2 expression was elevated in HNSC tissues, and ESCO2 depletion suppressed tumor cell migration in vitro and inhibited tumor growth in vitro and in vivo. Co-IP/MS and immunoblotting assays revealed the interaction between ESCO2 and STAT1 in HPC cells. STAT1-overexpression compromised ESCO2-mediated suppressive effects on HPC cell proliferation, viability, and migration. CONCLUSIONS: These findings suggest that ESCO2 is crucial in promoting HPC malignant progression through the STAT1 pathway and provides novel therapeutic targets for HPC treatment.


Assuntos
Segregação de Cromossomos , Neoplasias de Cabeça e Pescoço , Animais , Camundongos , Humanos , Camundongos Nus , Proliferação de Células , Carcinoma de Células Escamosas de Cabeça e Pescoço/genética , Linhagem Celular Tumoral , Fator de Transcrição STAT1/genética , Fator de Transcrição STAT1/metabolismo , Acetiltransferases/genética , Proteínas Cromossômicas não Histona/genética
4.
Analyst ; 148(20): 5124-5132, 2023 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-37681669

RESUMO

Targeted imaging is playing an increasingly important role in the early detection and precise diagnosis of cancer. This need has motivated research into sensory nanomaterials that can be constructed into imaging agents to serve as biosensors. Graphene quantum dots (GQDs) as a valuable nanoprobe show great potential for use in two-photon biological imaging. However, most as-prepared GQDs exhibit a low two-photon absorption cross-section, narrow spectral coverage, and "one-to-one" signal conversion mode, which greatly hamper their wide application in sensitive early-stage cancer detection. Herein, a versatile strategy has been employed to fabricate an aptamer Sgc8c-functionalized hybrid as a proof-of-concept of the signal amplification strategy for targeted cancer imaging. In this study, GQDs with two-photon imaging performance, and silica nanoparticles (SiO2 NPs) as nanocarriers to provide amplified recognition events by high loading of GQD signal tags, were adopted to construct a two-photon hybrid-based signal amplification strategy. Thus, the obtained hybrid (denoted SiO2@GQDs) enabled extremely strong fluorescence with a quantum yield up to 0.49, excellent photostability and biocompatibility, and enhanced bright two-photon fluorescence up to 2.7 times that of bare GQDs (excitation at 760 nm; emission at 512 nm). Moreover, further modification with aptamer Sgc8c showed little disruption to the structure of the SiO2@GQDs-hybrid and the corresponding two-photon emission. Hence, SiO2@GQDs-Sgc8c showed specific responses to target cells. Moreover, it could be used as a signal-amplifying two-photon nanoprobe for targeted cancer imaging with high specificity and great efficiency, which exhibits a distinct green fluorescence compared to that of GQDs-Sgc8c or SiO2@GQDs. This signal amplification strategy holds great potential for the accurate early diagnosis of tumors and offers new tools for the detection a wide variety of analytes in clinical application.


Assuntos
Grafite , Nanopartículas , Neoplasias , Pontos Quânticos , Humanos , Pontos Quânticos/química , Grafite/química , Dióxido de Silício/química , Nanopartículas/química , Oligonucleotídeos , Neoplasias/diagnóstico por imagem
5.
Artigo em Inglês | MEDLINE | ID: mdl-37944970

RESUMO

Objective: This study aims to provide clinical evidence for the treatment of idiopathic scoliosis (IS) by assessing the therapeutic effectiveness of combining functional rehabilitation training with orthosis. Methods: We enrolled a total of 94 IS patients who were admitted to our hospital from April 2020 to February 2022. These patients were randomly assigned into two groups: a research group (RG; n=47) receiving functional rehabilitation training combined with orthosis and a control group (CG; n=47) receiving orthosis treatment alone. Clinical outcomes were evaluated one year after treatment. We also measured the Cobb angle, apical vertebral rotation (AVR), and the distance between the vertical line of the sacrum and the spinous process of the scoliosis parietal vertebra before and after treatment to determine apical vertebral translation (AVT) from the sacral midline and lumbar range of motion (ROM). Patient quality of life was assessed using the Short-Form 36 Item Health Survey (SF-36). Results: After treatment, the research group exhibited significantly lower Cobb angles, AVR, and AVT, along with a higher overall response rate and greater lumbar ROM compared to the control group (P < .05). Post-treatment SF-36 scores increased in both groups, with notably higher scores in the research group (P < .05). Conclusions: Combining functional rehabilitation training with orthosis is an effective approach for the treatment of IS and holds substantial clinical significance.

6.
Exp Appl Acarol ; 89(3-4): 345-362, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-37027055

RESUMO

Transformer-2 (tra-2) is an important sex-determining gene in insects. It also plays a role in the reproduction of phytoseiid mites. We performed bioinformatic analyses for the tra-2 ortholog in Phytoseiulus persimilis (termed Pptra-2), measured its expression at different stages and quantitatively identified its function in reproduction. This gene encodes 288 amino acids with a conserved RRM domain. The peak of its expression was observed in adult females, especially ca. 5 days after mating. In addition, expression is also higher in eggs than in other stages and adult males. When Pptra-2 was silenced through RNA interference with oral delivery of dsRNA, 56% of the females had their egg hatching rates decreased in the first 5 days, from ca. 100% to ca. 20%, and maintained at low levels during the rest of the oviposition period. To detect other genes functionally related to Pptra-2, transcriptome analyses were performed on day 5 after mating. We compared mRNA expressions among interfered females with significantly reduced egg hatching rate, interfered females without significant hatching rate and CK. In total 403 differential genes were identified, of which 42 functional genes involved in the regulation of female reproduction and embryonic development were screened and discussed.


Assuntos
Ácaros , Reprodução , Masculino , Feminino , Animais , Ácaros/fisiologia , Oviposição , Interferência de RNA , Desenvolvimento Embrionário
7.
Plant Biotechnol J ; 20(8): 1487-1501, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35426230

RESUMO

Riboflavin is the precursor of essential cofactors for diverse metabolic processes. Unlike animals, plants can de novo produce riboflavin through an ancestrally conserved pathway, like bacteria and fungi. However, the mechanism by which riboflavin regulates seed development is poorly understood. Here, we report a novel maize (Zea mays L.) opaque mutant o18, which displays an increase in lysine accumulation, but impaired endosperm filling and embryo development. O18 encodes a rate-limiting bifunctional enzyme ZmRIBA1, targeted to plastid where to initiate riboflavin biosynthesis. Loss of function of O18 specifically disrupts respiratory complexes I and II, but also decreases SDH1 flavinylation, and in turn shifts the mitochondrial tricarboxylic acid (TCA) cycle to glycolysis. The deprivation of cellular energy leads to cell-cycle arrest at G1 and S phases in both mitosis and endoreduplication during endosperm development. The unexpected up-regulation of cell-cycle genes in o18 correlates with the increase of H3K4me3 levels, revealing a possible H3K4me-mediated epigenetic back-up mechanism for cell-cycle progression under unfavourable circumstances. Overexpression of O18 increases riboflavin production and confers osmotic tolerance. Altogether, our results substantiate a key role of riboflavin in coordinating cellular energy and cell cycle to modulate maize endosperm development.


Assuntos
Endosperma , Zea mays , Ciclo Celular/genética , Endosperma/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Proteínas de Plantas/genética , Riboflavina/genética , Riboflavina/metabolismo , Sementes , Zea mays/metabolismo
8.
Ophthalmologica ; 244(4): 326-333, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33045702

RESUMO

BACKGROUND: Retinoblastoma (RB) is a common intraocular malignant tumor in infants and young children. However, reports on the morphological descriptions of RB tumor cells from native and foreign scholars are rare. OBJECTIVES: To investigate the myelogram characteristics of RB with extraocular tumor extension and the morphological characteristics of tumor cells in the bone marrow and cerebrospinal fluid. METHODS: For the period from May 2011 to February 2015, we analyzed clinical data on 18 patients in our hospital diagnosed as having metastatic RB in the extraocular and other distant regions associated with clear bone marrow metastasis. The morphology of tumor cells in the bone marrow and cerebrospinal fluid was retrospectively analyzed after staining with Wright-Giemsa stain. A summary of the cytological characteristics was also presented. RESULTS: RB tumor cells in the bone marrow and cerebrospinal fluid not only appeared as aggregated clumps, but were distributed in a scattered manner. The tumor cells may present different characteristic morphologies in different cases, with different tumor cell smears from the same tumor mass even showing different features. According to the degree of tumor metastasis, changes in myelogram were significantly different. CONCLUSION: The tumor cells of RB patients show unique morphological characteristics in the bone marrow and cerebrospinal fluid. Therefore, correct identification of the cells is of great value in the diagnosis, staging, and prognosis of RB.


Assuntos
Neoplasias da Retina , Retinoblastoma , Medula Óssea , Pré-Escolar , Humanos , Lactente , Prognóstico , Neoplasias da Retina/diagnóstico , Retinoblastoma/diagnóstico , Estudos Retrospectivos
9.
J Dairy Sci ; 104(6): 6577-6587, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33741157

RESUMO

Bacillus cereus is one of the most important spoilage microorganisms in milk. The heat-resistant protease produced is the main factor that causes rotten, bitter off-flavors and age gelation during the shelf-life of milk. In this study, 55 strains of B. cereus were evaluated, of which 25 strains with protease production ability were used to investigate proteolytic activity and protease heat resistance. The results showed that B. cereus C58 had strong protease activity, and its protease also had the highest thermal stability after heat treatment of 70°C (30 min) and 100°C (10 min). The protease was identified as protease HhoA, with a molecular mass of 43.907 kDa. The protease activity of B. cereus C58 in UHT-sterilized whole milk (UHT milk) showed an increase with the growth of bacteria, especially during the logarithmic growth phase. In addition, the UHT milk incubated with protease from B. cereus C58 at 28°C (24 h) and 10°C (6 d) were used to evaluate the effects of protease on the quality of UHT milk, including protein hydrolysis and physical stability. The results showed that the hydrolysis of casein was κ-CN, ß-CN, and αS-CN successively, whereas whey protein was not hydrolyzed. The degree of protein hydrolysis, viscosity, and particle size of the UHT milk increased. The changes in protein and fat contents indicated that fat globules floated at 28°C and settled at 10°C, respectively. Meanwhile, confocal laser scanning microscopy images revealed that the protease caused the stability of UHT milk to decrease, thus forming age gelation.


Assuntos
Bacillus cereus , Leite , Animais , Temperatura Alta , Peptídeo Hidrolases , Temperatura
10.
J Dairy Sci ; 104(8): 8506-8516, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34053767

RESUMO

Effective testing tools for Escherichia coli O157:H7 can prevent outbreaks of foodborne illness. In this paper, a smartphone-based colorimetric aptasensor was developed using functionalized gold nanoparticles (GNP) and multi-walled carbon nanotubes (MWCNT) for monitoring E. coli O157:H7 in milk. The maximum absorption peak of GNP bonded with aptamer (Apt) generated evident transformation from 518 to 524 nm. The excess GNP-Apt was removed by functionalized MWCNT magnetized with carbonyl iron powder (CIP) and hybridized with a DNA probe, whereas the GNP-Apt immobilized on E. coli O157:H7 remained in the system. In the presence of a high-salt solution, the GNP-Apt that captured E. coli O157:H7 remained red, but the free GNP-Apt aggregated and appeared blue. The chromogenic results were analyzed by a smartphone-based colorimetric device that was fabricated using acrylic plates, a light-emitting diode, and a mobile power pack. To our knowledge, this was the first attempt to use a smartphone-based colorimetric aptasensor employing the capture of GNP-Apt coupled with separation of MWCNT@CIP probe to detect E. coli O157:H7. The aptasensor exhibited good reproducibility and no cross-reaction for other bacteria. A concentration of 8.43 × 103 cfu/mL of E. coli O157:H7 could be tested in pure culture, and 5.24 × 102 cfu/mL of E. coli O157:H7 could be detected in artificially contaminated milk after 1 h of incubation. Therefore, the smartphone-based colorimetric aptasensor was an efficient tool for the detection of E. coli O157:H7 in milk.


Assuntos
Técnicas Biossensoriais , Escherichia coli O157 , Nanopartículas Metálicas , Nanotubos de Carbono , Animais , Técnicas Biossensoriais/veterinária , Colorimetria/veterinária , Microbiologia de Alimentos , Ouro , Leite , Reprodutibilidade dos Testes , Smartphone
11.
J Dairy Sci ; 104(5): 5152-5165, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33663822

RESUMO

Due to the lack of specific genes for rapid detection methods of Cronobacter sakazakii in food samples, whole genome sequence analysis was performed in this investigation using the basic local alignment search tool. Forty-two DNA fragments unique to C. sakazakii were mined, then primers were designed and screened by PCR and loop-mediated isothermal amplification (LAMP). Two primer sets, CS1 and CS31, were found as specific and stable primers, with their corresponding nucleic acid targets the CSK29544_00235 gene and CSK29544_03484 gene, respectively. Furthermore, compared with 3 genes reported previously, these 2 genes were verified as more specific to C. sakazakii among Cronobacter species, by sequence similarity alignment using Cronobacter MLST databases (http://pubmlst.org/cronobacter). The specificity of the LAMP reaction approached 100% by using 48 bacterial strains, which included 22 C. sakazakii strains. Subsequently, LAMP was combined with visual lateral flow dipstick (LFD) based on the above 2 nucleic acid targets, and was demonstrated as a rapid, efficient method with high specificity. Finally, the detection sensitivity of this assay system for pure cultures and artificially contaminated milk was measured as 4.5 × 100 cfu/mL and 5.7 × 101 cfu/g, respectively. Total time to detection for this assay was within 2 h. Thus, the establishment of this LAMP-LFD method shows great significance and potential for rapid detection of C. sakazakii in powdered infant formula.


Assuntos
Cronobacter sakazakii , Cronobacter , Ácidos Nucleicos , Animais , Cronobacter/genética , Cronobacter sakazakii/genética , Microbiologia de Alimentos , Fórmulas Infantis , Técnicas de Diagnóstico Molecular , Tipagem de Sequências Multilocus/veterinária , Técnicas de Amplificação de Ácido Nucleico , Pós
12.
J Sci Food Agric ; 101(10): 4332-4341, 2021 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-33417239

RESUMO

BACKGROUND: Elemental selenium, as a new type of selenium supplement, can be prepared by microorganisms reducing inorganic selenium. In this study, Lactobacillus brevis JLD715 was incubated in broth containing different concentrations of sodium selenite (Na2 SeO3 ). RESULTS: The results showed that the bacterial biomass of L. brevis JLD715 decreased due to the inhibition of Na2 SeO3 . The cell membrane of L. brevis JLD715 treated with Na2 SeO3 was damaged, as evidenced by the reduction of intracellular ATP concentration, depolarization of cell membrane, reduction of intracellular pH and impairment of membrane integrity. In addition, we investigated the metabolism mechanism of Na2 SeO3 by L. brevis JLD715 based on transcriptome sequencing. A total of 461 genes were significantly differentially expressed under Na2 SeO3 treatment, of which 231 genes were up-regulated and 230 genes were down-regulated. These genes were involved in pathways such as pyruvate metabolism, fatty acid biosynthesis, selenocompound metabolism and nucleotide-binding oligomerization domain-like (NOD-like) receptor signaling. Meanwhile, the genes related to sulfhydryl oxidoreductase, electron carrier proteins and transmembrane transport proteins synthesis were significantly up-regulated. CONCLUSION: To sum up, the findings of this research will contribute to providing support for the application of L. brevis JLD715 in selenium-enriched functional foods. © 2021 Society of Chemical Industry.


Assuntos
Proteínas de Bactérias/genética , Levilactobacillus brevis/genética , Levilactobacillus brevis/metabolismo , Selenito de Sódio/metabolismo , Proteínas de Bactérias/metabolismo , Regulação Bacteriana da Expressão Gênica , Levilactobacillus brevis/crescimento & desenvolvimento , Transcriptoma
13.
J Cell Physiol ; 235(1): 380-393, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31232476

RESUMO

Atherosclerosis (AS), a chronic disorder of large arteries, is the underlying pathological process of heart disease and stroke. Former researchers have found that microRNAs (miRs) are involved in the several key processes of AS. Apolipoprotein E knockout (ApoE-/- ) mice fed a high-fat-diet (HFD) to establish AS model. The expression of miR-103 was characterized in the mice model. The effects of miR-103 on inflammation and endoplasmic reticulum stress (ERS) were analyzed when the expression of miR-103 was inhibited in ApoE -/- mice fed an HFD and human aortic endothelial cells (HAECs) exposed to oxidized low-density lipoprotein (ox-LDL). The relationship between miR-103 and phosphatase and tensin homolog (PTEN) was identified by luciferase activity detection and real-time quantitative polymerase chain reaction (RT-qPCR). Gain- and loss-function approaches were further applied for investigating the regulatory effects of miR-103 and PTEN on ERS. Role of MAPK signaling was then analyzed using PD98059 to block this pathway. miR-103 was highly expressed in the ApoEApoE -/- mice fed an HFD. Downregulation of miR-103 suppressed inflammation and ERS in endothelial cells isolated from ApoE -/- mice fed a HFD and ox-LDL-exposed HAECs. In addition, miR-103 can target PTEN and downregulate its expression. Overexpression of PTEN reversed the miR-103-induced activation of MAPK signaling. Moreover, PTEN upregulation or MAPK signaling inhibition ease miR-103-induced inflammation and ERS in vivo and in vitro. Thus, miR-103 depletion restrains the progression of AS through blocking PTEN-mediated MAPK signaling.


Assuntos
Aterosclerose/genética , Aterosclerose/patologia , Estresse do Retículo Endoplasmático/genética , MicroRNAs/genética , PTEN Fosfo-Hidrolase/metabolismo , Animais , Apolipoproteínas E/genética , Células Cultivadas , Modelos Animais de Doenças , Flavonoides/farmacologia , Células Endoteliais da Veia Umbilical Humana/patologia , Humanos , Inflamação/genética , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , MicroRNAs/antagonistas & inibidores
14.
J Cell Physiol ; 235(3): 2809-2824, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31535382

RESUMO

Breast cancer has become the most common cancer in women, and nontriple negative breast cancer (non-TNBC) accounts for 80-90% of all invasive breast cancers. Early detection, diagnosis, and treatment are considered key to a successful cure. Conventionally, breast imaging and needle core biopsy are used for detection and monitoring. However, small variations in volume might be ignored in imaging, and traditional biopsies are spatially and temporally limited, leading to a significant delay in cancer detection and thus prompting renewed focus on early and accurate diagnosis. In this article, we investigated whether there is an accurate molecule in peripheral blood that can help diagnose breast cancer. Similar to microRNAs, tRNA-derived fragments (tRFs) have been reported to be involved in many pathological processes in breast cancer, but whether they can serve as candidate biomarkers for breast cancer remains unclear. Using high-throughput sequencing technology, we identified 4,021 differentially expressed tRFs in normal and breast cancer cell lines, and eight tRFs were selected to establish a signature as a predictive biomarker of non-TNBC. Furthermore, quantitative reverse-transcriptase polymerase chain reaction was performed to verify the expression of the signature and analyze the correlation between dysregulated tRFs and breast cancer. The results indicated that tDR-7816, tDR-5334, and tDR-4733 might be promising biomarkers. Through further bioinformatics analysis, we predicted that tDR-7816 influences the xenobiotic metabolic processes that support the oncogenesis of breast cancer. In summary, our results provide a rationale for using circulating tDR-7816 expression as a novel potential biomarker for the diagnosis of patients with early non-TNBC.


Assuntos
Detecção Precoce de Câncer/métodos , RNA de Transferência/sangue , Neoplasias de Mama Triplo Negativas/diagnóstico , Neoplasias de Mama Triplo Negativas/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/genética , Carcinogênese/genética , Linhagem Celular Tumoral , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Células MCF-7 , Pessoa de Meia-Idade , RNA de Transferência/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa
15.
BMC Med Genet ; 21(1): 120, 2020 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-32487029

RESUMO

BACKGROUND: Familial benign chronic pemphigus, also known as Hailey-Hailey disease (HHD), is a clinically rare bullous Dermatosis. However the mechanism has not been clarified. The study aim to detect novel mutations in exons of ATP2C1 gene in HHD patients; to explore the possible mechnism of HHD pathogenesis by examining the expression profile of hSPCA1, miR-203, p63, Notch1 and HKII proteins in the skin lesions of HHD patients. METHODS: Genomic DNA was extracted from peripheral blood of HHD patients. All exons of ATP2C1 gene in HHD patients were amplified by PCR and the products were purified and sequenced. All related signaling proteins of interest were stained by using skin lesion tissues from HHD patients and miR-203 levels were also determined. RESULTS: One synonymous mutation c.G2598A (in exon 26), one nonsense mutation c.C635A and two missense mutations c.C1286A (p.A429D) and c. A1931G (p. D644G) were identified. The nonsense mutation changed codon UCG to stop codon UAG, causing a premature polypeptide chain of the functional region A. The two missense mutations were located in the region P (phosphorylation region) and the Mn binding site of hSPCA1. The level of hSPCA1 was significantly decreased in HHD patients compared to the normal human controls, accompanied by an increase of miR-203 level and a decrease of p63 and HKII levels. CONCLUSION: In our study, we found four mutations in HHD. Meanwhile we found increase of miR-203 level and a decrease of p63 and HKII levels. In addition, Notch1, which was negatively regulated p63, is downregulated. These factors may be involved in the signaling pathways of HHD pathogenesis. Our data showed that both p63 and miR-203 may have significant regulatory effects on Notch1 in the skin.


Assuntos
ATPases Transportadoras de Cálcio/genética , Estudos de Associação Genética , Predisposição Genética para Doença , Mutação , Pênfigo Familiar Benigno/diagnóstico , Pênfigo Familiar Benigno/genética , Sequência de Aminoácidos , Biópsia , ATPases Transportadoras de Cálcio/química , Éxons , Feminino , Estudos de Associação Genética/métodos , Humanos , Imuno-Histoquímica , Masculino , MicroRNAs/genética , Linhagem , Análise de Sequência de DNA , Transdução de Sinais , Pele/patologia
16.
J Dairy Sci ; 103(9): 7879-7889, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32600757

RESUMO

Escherichia coli O157:H7 is an extremely serious foodborne pathogen accounting for a vast number of hospitalizations. In this system, a simple, rapid, and safe compound method was developed based on carbonyl iron powder (CIP) and multiwalled carbon nanotubes (MWCNT). Then, the CIP@MWCNT-based aptasensor was constructed by strong π-stacking between nanocomposite and aptamer, single-strand DNA, causing fluorescent quenching of the dye-labeled aptamer. The restoration of dye fluorescence could be achieved when aptamer came off the surface of the CIP@MWCNT nanocomposite due to the presence of target bacteria. To the best of our knowledge, this fabrication of magnetic carbon nanotubes without irritating and corrosive reagents is described for the first time. The sensing platform was also an improvement on the conventional formation of the aptasensor between carbon materials and DNA aptamer. The nanocomposite was verified by diverse characterization of zeta potential, Fourier-transform infrared spectroscopy, transmission electron microscopy, and energy dispersive x-ray analysis. The CIP@MWCNT-based aptasensor was an effective nanoplatform for quantitative detection of E. coli O157:H7, and was measured to have high specificity, good reproducibility, and strong stability. The aptasensor's capacity to quantify E. coli O157:H7 was as low as 7.15 × 103 cfu/mL in pure culture. The detection limit of E. coli O157:H7 was 3.15 × 102 cfu/mL in contaminated milk after 1 h of pre-incubation. Hence, the developed assay is a new possibility for effective synthesis of nanocomposites and sensitive tests of foodborne pathogens in the dairy industry.


Assuntos
Escherichia coli O157/isolamento & purificação , Fluorometria/veterinária , Leite/microbiologia , Nanocompostos , Animais , Técnicas Biossensoriais , Fluorometria/métodos , Microbiologia de Alimentos , Nanotubos de Carbono , Reprodutibilidade dos Testes
17.
J Cell Biochem ; 120(3): 3801-3812, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30506709

RESUMO

Benign chronic familial pemphigus or Hailey-Hailey disease (HHD, OMIM 169600) is a rare, autosomal dominant blistering skin disorder characterized by suprabasal cell separation (acantholysis) of the epidermis. To date, the proteomic changes in skin lesions from HHD patients has not been reported yet. In this study, a sample of skin lesions from HHD patients was collected for isobaric tags for relative and absolute quantitation to analyze proteome changes compared with unaffected individuals. The 134 differentially expressed proteins were assigned to at least one Gene Ontology term, and 123 annotated proteins with significant matches were assigned to 187 known metabolic or signaling pathways listed in the Kyoto Encyclopedia of Genes and Genomes. Most of the altered proteins in skin lesions of HHD patients were enriched in pathways involved in the PI3K-Akt signaling, focal adhesion, extracellular matrix (ECM)-receptor interaction, and protein digestion and absorption, such as collagen family members, microfibril-associated glycoprotein 4 and plakophilin. The changes of proteins related to cell adhesion, ECM-receptor interaction, and protein folding and glycosylation suggested that strategy targeted to alter cell junction and extracellular microenvironment might provide a potential treatment for HHD.


Assuntos
Matriz Extracelular/genética , Adesões Focais/genética , Pênfigo Familiar Benigno/genética , Proteoma/genética , Receptores de Superfície Celular/genética , Adulto , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Estudos de Casos e Controles , Colágeno/genética , Colágeno/metabolismo , Enciclopédias como Assunto , Epiderme/metabolismo , Epiderme/patologia , Matriz Extracelular/patologia , Proteínas da Matriz Extracelular/genética , Proteínas da Matriz Extracelular/metabolismo , Feminino , Adesões Focais/metabolismo , Adesões Focais/patologia , Ontologia Genética , Glicoproteínas/genética , Glicoproteínas/metabolismo , Humanos , Queratinócitos/metabolismo , Queratinócitos/patologia , Redes e Vias Metabólicas/genética , Anotação de Sequência Molecular , Pênfigo Familiar Benigno/metabolismo , Pênfigo Familiar Benigno/patologia , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , Placofilinas/genética , Placofilinas/metabolismo , Mapeamento de Interação de Proteínas , Proteoma/metabolismo , Proteômica/métodos , Proteínas Proto-Oncogênicas c-akt/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptores de Superfície Celular/metabolismo , Transdução de Sinais
18.
Environ Health Prev Med ; 22(1): 45, 2017 May 11.
Artigo em Inglês | MEDLINE | ID: mdl-29165156

RESUMO

BACKGROUND: It is generally acknowledged that the determination of harmful chemical compounds excreted into saliva is useful for assessing their exposure levels. The aim of the present study was to compare the total arsenic and its species in saliva and urine samples collected from the people residing in an arsenic-contaminated area of China and to further verify the feasibility of using salivary arsenic as a new biomarker of arsenic exposure. METHODS: Total arsenic and speciation analyses in urine and saliva samples among 70 residents exposed to arsenic from drinking water in Shanxi, China were carried out by high-performance liquid chromatography-inductively coupled plasma-mass spectrometry (HPLC-ICP/MS). RESULTS: The result showed that, total arsenic concentration in saliva was relatively lower than in urine samples, but it existed a strong positive correlation with total urinary arsenic, drinking water arsenic and different skin lesions. For arsenic metabolism analyses, AsIII, AsV, MMA, and DMA were detected in all of the urine samples with the dominating species of DMA (73.2%). Different with urinary arsenic species, most arsenic species in saliva were not methylated. The major species in saliva was iAs (AsIII + AsV, 76.18%), followed by DMA (13.08%) and MMA (9.13%). And the primary methylation index (PMI), second methylation index (SMI) and proportion of the four different species (AsIII, AsV, MMA, and DMA) in saliva showed no significant positive relationship with that of in urine. CONCLUSIONS: These findings indicated saliva may be used as a useful tool for biological monitoring of total arsenic exposure in the crowd rather than an efficient tool for assessing arsenic metabolism in human body after exposed to arsenic.


Assuntos
Arsênio/metabolismo , Arsenicais/metabolismo , Poluentes Químicos da Água/metabolismo , Adulto , Idoso , Arsênio/urina , Arsenicais/urina , Biomarcadores/metabolismo , Biomarcadores/urina , China , Cromatografia Líquida de Alta Pressão , Água Potável/análise , Monitoramento Ambiental , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Saliva/química , Poluentes Químicos da Água/urina , Adulto Jovem
19.
Beijing Da Xue Xue Bao Yi Xue Ban ; 46(3): 424-8, 2014 Jun 18.
Artigo em Zh | MEDLINE | ID: mdl-24943022

RESUMO

OBJECTIVE: To establish the control charts for early warning of diarrhea based on the syndromic surveillance data from enteric clinic in Beijing. METHODS: The outpatient data from enteric clinic of a Grade Three General hospital in Haidian district, Beijing from April 1 to Oct. 31, 2009 and from May 1 to Nov.10, 2010 were collected, according to the moving average method, the baseline calculated, the value of probability α and µα, the early warning value based on the formula "w=Xj+µαSj" calculated and the early warning control charts drew at last. RESULTS: According to the harmfulness, the severity and controllability of diarrheal diseases, the value of probability α was determined as 0.01, then µα (unilateral) as 2, based on the early warning value, the control charts of diarrheal diseases, bacillary dysentery and other infectious diarrhea were established. CONCLUSION: The enteric clinic requires to further collect baseline data to evaluate and continuously adjust the established control charts for the best early warning model in accordance with the enteric clinic.


Assuntos
Diarreia , Vigilância da População/métodos , Interpretação Estatística de Dados , Notificação de Doenças , Disenteria Bacilar , Humanos , Pacientes Ambulatoriais
20.
Talanta ; 269: 125457, 2024 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-38039678

RESUMO

Escherichia coli O157: H7 (E. coli O157: H7) is one of the most common foodborne pathogens and is widespread in food and the environment. Thus, it is significant for rapidly detecting E. coli O157: H7. In this study, a colorimetric aptasensor based on aptamer-functionalized magnetic beads, exonuclease III (Exo III), and G-triplex/hemin was proposed for the detection of E. coli O157: H7. The functional hairpin HP was designed in the system, which includes two parts of a stem containing the G-triplex sequence and a tail complementary to cDNA. E. coli O157: H7 competed to bind the aptamer (Apt) in the Apt-cDNA complex to obtain cDNA. The cDNA then bound to the tail of HP to trigger Exo III digestion and release the single-stranded DNA containing the G-triplex sequence. G-triplex/hemin DNAzyme could catalyze TMB to produce visible color changes and detectable absorbance signals in the presence of H2O2. Based on the optimal conditions, E. coli O157: H7 could be detected down to 1.3 × 103 CFU/mL, with a wide linear range from 1.3 × 103 to 1.3 × 107 CFU/mL. This method had a distinguished ability to non-target bacteria, which showed good specificity. In addition, the system was successfully applied to detect E. coli O157: H7 in milk samples.


Assuntos
Aptâmeros de Nucleotídeos , DNA Catalítico , Escherichia coli O157 , Escherichia coli O157/genética , Hemina , Colorimetria/métodos , DNA Complementar , Peróxido de Hidrogênio , Aptâmeros de Nucleotídeos/genética , Fenômenos Magnéticos , Microbiologia de Alimentos
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