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1.
Brain ; 2024 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-38875478

RESUMO

USP25 encodes ubiquitin-specific proteases 25, a key member of deubiquitinating enzyme family and is involved in neural fate determination. Although abnormal expression in Down's syndrome was reported previously, the specific role of USP25 in human diseases has not been defined. In this study, we performed trio-based whole exome sequencing in a cohort of 319 cases (families) with generalized epilepsy of unknown etiology. Five heterozygous USP25 variants including two de novo and three co-segregated variants were determined in eight individuals affected by generalized seizures and/or febrile seizures from five unrelated families. The frequency of USP25 variants showed a significantly high aggregation in this cohort compared to the East Asian population and all populations in the gnomAD database. The mean onset ages of febrile and afebrile seizures were 10 months (infancy) and 11.8 years (juvenile), respectively. The patients achieved seizure freedom except one had occasional nocturnal seizures at the last follow-up. Two patients exhibited intellectual disability. Usp25 was ubiquitously expressed in mouse brain with two peaks on embryonic days (E14‒E16) and postnatal day 21, respectively. Similarly, USP25 expressed in fetus/early childhood stage with a second peak at approximately 12‒20 years old in human brain, consistent with the seizure onset age at infancy and juvenile in the patients. To investigate the functional impact of USP25 deficiency in vivo, we established Usp25 knock-out mice, which showed increased seizure susceptibility compared to wild-type mice in pentylenetetrazol-induced seizure test. To explore the impact of USP25 variants, we employed multiple functional detections. In HEK293T cells, the severe phenotype associated variant (p.Gln889Ter) led to a significant reduction of mRNA and protein expressions but formed a stable truncated dimers with increment of deubiquitinating enzyme activities and abnormal cellular aggregations, indicating a gain-of-function effect. The p.Gln889Ter and p.Leu1045del increased neuronal excitability in mice brain, with a higher firing ability in p.Gln889Ter. These functional impairments align with the severity of the observed phenotypes, suggesting a genotype-phenotype correlation. Hence, a moderate association between USP25 and epilepsy was noted, indicating USP25 is potentially a predisposing gene for epilepsy. Our results from Usp25 null mice and the patient-derived variants indicated that USP25 would play epileptogenic role via loss-of-function or gain-of-function effects. The truncated variant p.Gln889Ter would have profoundly different effect on epilepsy. Together, our results underscore the significance of USP25 heterozygous variants in epilepsy, thereby highlighting the critical role of USP25 in the brain.

2.
Anal Chem ; 96(12): 5046-5055, 2024 Mar 26.
Artigo em Inglês | MEDLINE | ID: mdl-38488055

RESUMO

Bimodal-type multiplexed immunoassays with complementary mode-based correlation analysis are gaining increasing attention for enhancing the practicability of the lateral flow immunoassay (LFIA). Nonetheless, the restriction in visually indistinguishable multitargets induced by a single fluorescent color and difficulty in single acceptor ineffectual fluorescence quenching due to the various spectra of multiple different donors impede the further execution of colorimetric-fluorescence bimodal-type multiplexed LFIAs. Herein, the precise spectral overlap-based donor-acceptor pair construction strategy is proposed by regulating the size of the nanocore, coating it with an appropriate nanoshell, and selecting a suitable fluorescence donor with distinct colors. By in situ coating Prussian blue nanoparticles (PBNPs) on AuNPs with a tunable size and absorption spectrum, the resultant APNPs demonstrate efficient fluorescence quenching ability, higher colloidal stability, remarkable colorimetric intensity, and an enhanced antibody coupling efficiency, all of which facilitate highly sensitive bimodal-type LFIA analysis. Following integration with competitive-type immunoreaction, this precise spectral overlap-supported spatial separation traffic light-typed colorimetric-fluorescence dual-response assay (coined as the STCFD assay) with the limits of detection of 0.013 and 0.152 ng mL-1 for ractopamine and clenbuterol, respectively, was proposed. This work illustrates the superiority of the rational design of a precise spectral overlap-based donor-acceptor pair, hinting at the enormous potential of the STCFD assay in the point-of-care field.


Assuntos
Clembuterol , Nanopartículas Metálicas , Ouro , Imunoensaio , Fenômenos Químicos , Limite de Detecção
3.
Opt Express ; 32(8): 13552-13561, 2024 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-38859322

RESUMO

The effect of crystal-water contents on the optical properties and dielectric characteristics of calcium sulfate in the THz band is investigated. The complex dielectric constant and conductivity are analyzed using the Drude-Smith model. The refractive index and absorption coefficient are linearly increased with the content of crystal-water, and the corresponding linear fitting lines of R2 over 0.97 are obtained. The dielectric properties of calcium sulfate are significantly affected by the crystal-water content. These results indicate that a new method to quantitative measurement of the crystal-water content in hydrous minerals is provided.

4.
FASEB J ; 37(12): e23285, 2023 12.
Artigo em Inglês | MEDLINE | ID: mdl-37933950

RESUMO

Although certain progress has been made in treating canine inflammatory bowel disease (IBD), a large proportion of dogs have a poor prognosis and may develop resistance and side effects. Therefore, it is of great significance to prevent or alleviate canine IBD through nutritional intervention. Plant polyphenol can interact with intestinal bacteria and has important prospects in the intestinal health improvement. This study evaluated the effect of grape seed proanthocyanidin (GSP), a plant-derived natural polyphenol, on Labrador Retrievers with mild IBD. In Experiment 1 of this study, GSP alleviated persistent intestinal inflammation in canines by improving inflammatory indexes and reducing intestinal permeability. Moreover, GSP treatment increased the abundance of bacteria with potential anti-inflammatory properties and engaging bile acid metabolism, including Ruminococcaceae, Faecalibacterium, Ruminococcus_torques_group, and Lachnospiraceae_NK4A136_group. Notably, targeted metabolomic analysis identified elevated productions of fecal chenodeoxycholic acid and its microbial transformation product lithocholic acid, which might contribute to relieving canine intestinal inflammation. Further, in Experiment 2, fecal microbiota transplantation was used to determine whether gut microbiota is a potential mechanism for GSP efficacy. Dogs with mild IBD received the fecal microbiota from the group administered GSP and mirrored the improvement effects of GSP, which results verified that gut microbial alteration could be an underlying mechanism for GSP efficiency on canine IBD. Our findings highlight that the mechanism of the GSP function on canine IBD is mediated by altering gut microbial composition and improving bile acid metabolism. This study proposes a natural polyphenol-based dietary strategy for improving canine intestinal health.


Assuntos
Microbioma Gastrointestinal , Doenças Inflamatórias Intestinais , Cães , Animais , Ácidos e Sais Biliares , Doenças Inflamatórias Intestinais/microbiologia , Inflamação , Polifenóis/farmacologia
5.
Appl Opt ; 63(16): 4251, 2024 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-38856600

RESUMO

This publisher's note serves to correct errors in Appl. Opt.63, 2528 (2024)APOPAI0003-693510.1364/AO.517400.

6.
Appl Opt ; 63(10): 2528-2534, 2024 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-38568532

RESUMO

Terahertz time-domain spectroscopy was first used to establish a correlation with the whole-rock iron (TFe) content in different depths of the Bayan Obo protolith. Compared with element content obtained by the traditional method of X-ray fluorescence spectroscopy (XRF), a similar tendency of the absorption coefficient and refractive index is presented. Furthermore, three machine learning algorithms, namely, partial least squares regression (PLSR), random forest (RF), and multi-layer perceptron (MLP), were used to develop a quantitative analytical model for TFe content of the protolith minerals. Among the three algorithms, MLP has the highest detection accuracy, with a model coefficient of determination R 2 reaching up to 0.945. These findings demonstrate that terahertz time-domain spectroscopy can be used to rapidly quantify the TFe elemental content of protolith, providing a method of detecting the content of mineral components.

7.
Small ; 19(43): e2301598, 2023 10.
Artigo em Inglês | MEDLINE | ID: mdl-37381671

RESUMO

Engineered collaborative size regulation and shape engineering of multi-functional nanomaterials (NPs) offer extraordinary opportunities for improving the analysis performance. It is anticipated to address the difficulty in distinguishing color changes caused by subtle variations in target concentrations, thereby facilitating the highly sensitive analysis of lateral flow immunoassays (LFIAs). Herein, tremella-like gold-manganese oxide (Au-MnOx ) nanoparticles with precise MnCl2 regulation are synthesized as immuno signal tracers via a facile one-step redox reaction in alkaline condition at ambient temperature. Avail of the tunable elemental composition and anisotropy in morphology, black-colored tremella-like Au-MnOx exhibits superb colorimetric signal brightness, enhanced antibody coupling efficiency, marvelous photothermal performance, and unrestricted immunological recognition affinity, all of which facilitate highly sensitive multi-signal transduction patterns. In conjunction with the handheld thermal reader device, a bimodal-type LFIA that combines size-regulation- and shape-engineering-mediated colorimetric-photothermal dual-response assay (coined as the SSCPD assay) with a limit of detection of 0.012 ng mL-1 for ractopamine (RAC) monitoring is achieved by integrating Au-MnOx with the competitive-type immunoreaction. This work illustrates the effectiveness of this strategy for establishing high-performance sensing, and the SSCPD assay may be extended to a wide spectrum of future point-of-care (POC) diagnostic applications.


Assuntos
Nanopartículas Metálicas , Nanopartículas , Ouro , Imunoensaio , Anticorpos , Colorimetria , Limite de Detecção
8.
Opt Lett ; 48(24): 6500-6503, 2023 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-38099783

RESUMO

We propose and experimentally demonstrate a secure quantum noise stream cipher transmission system that integrates key generation and distribution. At the stage of carrier phase recovery, the estimated phase noise is used to generate randomness keys without additional equipment. Based on direct sequence spread spectrum technology, we integrate the distributed keys with quantum noise stream cipher signals. The key distribution and encryption transmission can be completed simultaneously without occupying additional bandwidth or time slots. By changing the position of distributed keys in the encryption base, the BER performance of QAM/QNSC signals cannot be affected by the keys. Experimental results demonstrate that the 54.5 Mbps key distribution and 31 Gbps encryption transmission without OSNR penalty can be achieved simultaneously over a 120 km standard single-mode fiber.

9.
Respir Res ; 24(1): 296, 2023 Nov 25.
Artigo em Inglês | MEDLINE | ID: mdl-38007420

RESUMO

Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive scarring interstitial lung disease with an unknown cause. Some patients may experience acute exacerbations (AE), which result in severe lung damage visible on imaging or through examination of tissue samples, often leading to high mortality rates. However, the etiology and pathogenesis of AE-IPF remain unclear. AE-IPF patients exhibit diffuse lung damage, apoptosis of type II alveolar epithelial cells, and an excessive inflammatory response. Establishing a reliable animal model of AE is critical for investigating the pathogenesis. Recent studies have reported a variety of animal models for AE-IPF, each with its own advantages and disadvantages. These models are usually established in mice with bleomycin-induced pulmonary fibrosis, using viruses, bacteria, small peptides, or specific drugs. In this review, we present an overview of different AE models, hoping to provide a useful resource for exploring the mechanisms and targeted therapies for AE-IPF.


Assuntos
Fibrose Pulmonar Idiopática , Humanos , Animais , Camundongos , Fibrose Pulmonar Idiopática/diagnóstico , Pulmão , Modelos Animais , Progressão da Doença
10.
BMC Infect Dis ; 23(1): 678, 2023 Oct 11.
Artigo em Inglês | MEDLINE | ID: mdl-37821800

RESUMO

BACKGROUND: COVID-19 has been a public health emergency of international concern (PHEIC) for a lengthy period of time. The novel coronavirus is primarily spread via aerosols at a short distance, with infected individuals releasing large amounts of aerosols when speaking and coughing. However, there is an open question regarding whether mouthwash could effectively reduce virus transmission during the COVID-19 pandemic and support the prevention of infection among medical workers. METHODS: Cochrane Library, PubMed, Web of Science, and Embase databases were systematically searched from the inception of each database to January 12, 2023 for currently available randomized clinical trials (RCTs) on the effect of mouthwash on novel coronavirus load in the oral cavity in COVID-19 patients. The treatment group received mouthwash for rinsing the mouth, while the control group received a placebo or distilled water for COVID-19 patients. The primary outcomes were CT value and viral load. Odds ratios (ORs) were estimated using a random-effects model. Subgroup and sensitivity analyses were performed to minimize the bias and the impact of heterogeneity. RESULTS: Thirteen RCTs were included. Seven studies reported the intervention effect of mouthwash on the CT value of novel coronavirus. The analysis results showed that the mouthwash group had a positive impact on the CT value of novel coronavirus [ SMD = 0.35, 95% CI (0.21, 0.50)] compared with the control group. In addition, subgroup analysis showed a significant positive effect of mouthwash on CT values in the treatment group compared with the control group, with chlorhexidine (CHX) [SMD = 0.33, 95% CI (0.10, 0.56)], povidone-iodine (PVP-I) [SMD = 0.61, 95% CI (0.23, 0.99)], or hydrogen peroxide (HP) [SMD = 1.04, 95% CI (0.30, 1.78)] as an ingredient of the mouthwash. Six studies reported the intervention effect of mouthwash on the viral load, 263 cases in the treatment group and 164 cases in the control group. The analysis results showed that there was no statistical difference between the mouthwash group and the control group in the viral load of novel coronavirus [SMD = -0.06, 95% CI (-0.18, 0.05)]. In the subgroup analysis by measurement time, there were statistically significant differences between the mouthwash and control groups for CT values [SMD = 0.52, 95% CI (0.31, 0.72)] and viral load [SMD = - 0.32, 95% CI (- 0.56, - 0.07)] within 30 min of gargling. CONCLUSIONS: In summary, mouthwash has some efficacy in reducing the viral load of novel coronavirus, especially within 30 min after rinsing the mouth. Mouthwash containing CHX, PVP-I and HP all had significant positive effects on CT values, and PVP-I-containing mouthwash may be a promising option to control novel coronavirus infections and relieve virus-related symptoms. However, studies on the dose and frequency of use of mouthwash for infection control are still lacking, which may limit the clinical application of mouthwash. TRIAL REGISTRATION: Protocol registration: The protocol was registered at PROSPERO (CRD42023401961).


Assuntos
COVID-19 , Antissépticos Bucais , Humanos , Antissépticos Bucais/uso terapêutico , Antissépticos Bucais/efeitos adversos , SARS-CoV-2 , Povidona-Iodo , Carga Viral , Aerossóis e Gotículas Respiratórios , Clorexidina/uso terapêutico , Peróxido de Hidrogênio
11.
Cell Mol Biol (Noisy-le-grand) ; 69(1): 13-18, 2023 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-37213163

RESUMO

Sertoli cells, the only somatic cells in testis seminiferous tubules, provide a supporting microenvironment for male germ cells and play essential roles in spermatogenesis. The insulin-degrading enzyme (IDE), a ubiquitous zinc peptidase of the inverzincin family, plays crucial role in sperm production, as IDE-knockout mice presented decreased testis weight and impaired sperm viability and morphology. However, whether and how IDE affects swine Sertoli cell proliferation remains unclear. Thus, in the present study, we aimed to evaluate the effects of IDE on the proliferation of swine Sertoli cells, as well as its underlying molecular mechanism. After knocking down IDE expression with small interfering RNA transfection, we analyzed the proliferation of swine Sertoli cells as well as the expression of related regulatory factors (WT1, ERK, and AKT). The results showed that IDE knockdown promoted swine Sertoli cell proliferation and increased WT1 expression, possibly through activating ERK and AKT. Overall, our findings suggest that IDE may be involved in male reproduction by regulating Sertoli cell proliferation, which provides new information to better understand the regulatory mechanisms of swine Sertoli cells and improve the reproductive traits of male pigs.


Assuntos
Insulisina , Células de Sertoli , Animais , Masculino , Proliferação de Células , Insulisina/genética , Insulisina/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Sêmen , Células de Sertoli/metabolismo , Suínos , Testículo/metabolismo
12.
Sens Actuators B Chem ; 390: 133950, 2023 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-37193119

RESUMO

Mpox (formerly referred as Monkeypox) outbreak has been declared a Public Health Emergency of International Concern. However, traditional polymerase chain reaction (PCR) diagnostic technology is not ideal for on-site applications. To conduct the sample-to-result Mpox viral particles detection outside the laboratories, we developed an easy-to-operate palm-size pouch, termed Mpox At-home Self-Test and point-of-caRe Pouch (MASTR Pouch). In this MASTR Pouch, the fast and accurate visualization was achieved by incorporating recombinase polymerase amplification (RPA) with clustered regularly interspaced short palindromic repeat (CRISPR)/Cas12a system. From viral particle lysis to naked eye readout, MASTR Pouch required only four simple steps to accomplish the analysis process within 35 min. Fifty-three Mpox pseudo-viral particles in exudate (10.6 particles/µL) were able to be detected. To verify the practicability, 104 mock Mpox clinical exudate specimens were tested. The clinical sensitivities were determined to be 91.7%- 95.8%. There was no false-positive result, validating the 100% clinical specificity. MASTR Pouch approaches the WHO's ASSURD criteria for point-of-care diagnostic, which will be beneficial for mitigating Mpox's global spread. The versatility potential of MASTR Pouch could further revolutionize infection diagnosis.

13.
Opt Express ; 30(22): 40645-40656, 2022 Oct 24.
Artigo em Inglês | MEDLINE | ID: mdl-36298995

RESUMO

Quantum noise stream cipher where encrypted signals are masked by quantum noise and ASE noise provides a physical layer of security. It requires the transmitter and the receiver to share a stream cipher that is generated from a PRNG. Yet a correlation attack threatens its security due to the mathematical properties of PRNG. This paper discusses the security of QNSC system under correlation attacks. Our experiment results find that the security of the whole system depends on the cycle to refresh the seed key and the correlation between the incepted running key, original running key, and seed key. Furthermore, it is important to provide security for the QNSC system by maintaining low optical power. Besides, this new analytical method provides quantitative security analysis for a QNSC system under a correlation attack.

14.
Mikrochim Acta ; 189(6): 218, 2022 05 11.
Artigo em Inglês | MEDLINE | ID: mdl-35546375

RESUMO

A dual-signal photometric/fluorometric assay was established for rapid, qualitative, and quantitative detection of Salmonella typhimurium (S. typhimurium). This method was composed of two parts: (1) a single-step photometric (SSC) assay containing gold nanoparticles (AuNPs), poly-diallyldimethylammonium chloride (PDDA), and S. typhimurium-specific aptamer, and (2) a fluorescence (FL) assay containing carboxyl-modified CdSe/ZnS quantum dots (QDs-COOH). Users just need to drop samples contaminated with S. typhimurium into SSC assay; the apparent color change from red to blue can be observed in a short time (20 min). A smartphone app was developed to read the semiquantitative result. By subsequently adding one drop of FL assay into the reaction mixture, the generated fluorescence intensity reflected the concentration of S. typhimurium. The naked eye limit of detection (LOD) and fluorescent LOD were 103 cfu/mL and 10 cfu/mL, respectively. This method exhibited good selectivity. The reliability and practicability were verified by testing contaminated food, drinking water, and pets' urine.


Assuntos
Ouro , Nanopartículas Metálicas , Limite de Detecção , Reprodutibilidade dos Testes , Salmonella typhimurium
15.
Int J Mol Sci ; 23(23)2022 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-36499554

RESUMO

A food allergy is caused by an abnormal immune reaction and can induce serious intestinal inflammation and tissue damage. Currently, the avoidance of food allergens is still the most effective way to prevent or reduce allergic symptoms, so the development of new strategies to treat allergies is important. Avenanthramide (AVA) is a bioactive polyphenol derived from oats with a wide range of biological activities; however, it is still not clear whether or how AVA alleviates intestinal damage under allergic situations. The aim of this study was to explore the effect of AVA on the small intestinal damage in an ovalbumin (OVA)-induced food allergy model and its mechanism. In experiment 1, 10 mg/kg bw and 20 mg/kg bw doses of AVA both decreased the serum levels of OVA-specific IgE, histamine, and prostaglandin D induced by OVA. The AVA administration relieved inflammation indicated by the lower serum concentrations of pro-inflammatory cytokines including interleukin-1ß, IL-6, and tumor necrosis factor-α. The levels of tight junction proteins including Claudin-1, ZO-1, and Occludin in the jejunum were elevated after AVA administration, accompanied by the improved intestinal morphology. Furthermore, AVA elevated the protein expression of heat shock protein 70 (Hsp70) and inhibited the phosphorylation of nuclear factor kappa-B (NF-κB), thus the apoptozole, which a Hsp70 inhibitor, was applied in experiment 2 to assess the contribution of Hsp70-NF-κB signaling to the effects of AVA. In the experiment 2, the inhibition of Hsp70 signaling treatment abolished the beneficial effects of AVA on the small intestinal damage and other allergic symptoms in mice challenged with OVA. Taken together, our results indicated that AVA exerted an intestinal protection role in the OVA-induced allergy, the mechanism of which was partly mediated by the Hsp70-NF-κB signaling.


Assuntos
Hipersensibilidade Alimentar , Intestino Delgado , NF-kappa B , Animais , Camundongos , Citocinas/metabolismo , Hipersensibilidade Alimentar/tratamento farmacológico , Proteínas de Choque Térmico HSP70/metabolismo , NF-kappa B/metabolismo , Ovalbumina/farmacologia , Transdução de Sinais , Intestino Delgado/metabolismo
16.
Sensors (Basel) ; 21(10)2021 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-34065696

RESUMO

Semiconductor (SC)-based field-effect transistors (FETs) have been demonstrated as amazing enhancer gadgets due to their delicate interface towards surface adsorption. This leads to their application as sensors and biosensors. Additionally, the semiconductor material has enormous recognizable fixation extends, high affectability, high consistency for solid detecting, and the ability to coordinate with other microfluidic gatherings. This review focused on current progress on the semiconductor-interfaced FET biosensor through the fundamental interface structure of sensor design, including inorganic semiconductor/aqueous interface, photoelectrochemical interface, nano-optical interface, and metal-assisted interface. The works that also point to a further advancement for the trademark properties mentioned have been reviewed here. The emergence of research on the organic semiconductor interface, integrated biosensors with Complementary metal-oxide-semiconductor (CMOS)-compatible, metal-organic frameworks, has accelerated the practical application of biosensors. Through a solid request for research along with sensor application, it will have the option to move forward the innovative sensor with the extraordinary semiconductor interface structure.


Assuntos
Técnicas Biossensoriais , Transistores Eletrônicos , Metais , Óxidos , Semicondutores
17.
Int J Mol Sci ; 22(22)2021 Nov 11.
Artigo em Inglês | MEDLINE | ID: mdl-34830091

RESUMO

Apigenin, a common dietary flavonoid abundantly present in a variety of fruits and vegetables, has promising anticancer properties. As an effector of apigenin in myoblasts, protein arginine methyltransferase 7 (Prmt7) is required for male germ cell development. However, whether apigenin may influence male reproductive health through Prmt7 is still unclear. To this end, mouse spermatogonia were treated with different concentrations (2.5 to 50 µM) of apigenin for 48 h, which showed that apigenin could cause reduced cell proliferation in conjunction with longer S phase and G2/M phase (with concentrations of 10 and 20 µM, respectively), and increased apoptosis of spermatogonia (with concentration of 20 µM). Reduced Prmt7 expression was found in 20 µM apigenin-treated spermatogonia. Moreover, siRNA-induced Prmt7 knockdown exhibited similar influence on spermatogonia as that of apigenin treatment. In mechanistic terms, transcriptome analysis revealed 287 differentially expressed genes between Prmt7-downregulated and control spermatogonia. Furthermore, rescue experiments suggested that the effects of apigenin on spermatogonia might be mediated through the Prmt7/Akt3 pathway. Overall, our study supports that apigenin can interfere with mouse spermatogonial proliferation by way of the downregulated Prmt7/Akt3 pathway, which demonstrates that the concentration should be taken into account in future applications of apigenin for cancer therapy of men.


Assuntos
Apigenina/farmacologia , Proliferação de Células/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Proteína-Arginina N-Metiltransferases/biossíntese , Proteínas Proto-Oncogênicas c-akt/biossíntese , Transdução de Sinais/efeitos dos fármacos , Espermatogônias/metabolismo , Animais , Masculino , Camundongos , Camundongos Endogâmicos ICR , Saúde Reprodutiva
18.
Med Mycol ; 58(1): 61-65, 2020 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-30927430

RESUMO

Sporothrix globosa is an important clinical pathogen in the Sporothrix complex, which is causing sporotrichosis. S. globosa is distributed worldwide, especially in Asia. The transmission medium of S. globosa is mainly contaminated soil or decaying vegetation, and the infection usually caused by transcutaneous trauma, through which the fungal conidia or yeast cells enter the host. Although the clinical manifestations of sporotrichosis caused by S. globosa is always benign, there have been several outbreaks worldwide. In this study, we established a novel real-time polymerase chain reaction (PCR) method based on the internal transcribed spacer (ITS) sequence for the identification of S. globosa. The assay was further evaluated by clinical specimens obtained from patients of sporotrichosis. The sensitivity and specificity of the real-time PCR method was both 100%. The detection limit was 10 fg. The positive detection rate for 30 clinical specimens, which were confirmed infected by S. globosa, was 100%. The real-time PCR method established in this paper is a rapid, sensitive and specific method for the identification of S. globosa. It can detect S. globosa in clinical specimen from patients with sporotrichosis, which is helpful for fast clinical diagnosis.


Assuntos
Reação em Cadeia da Polimerase em Tempo Real/métodos , Sporothrix/genética , Esporotricose/diagnóstico , Primers do DNA/genética , DNA Fúngico/genética , DNA Intergênico/genética , Humanos , Limite de Detecção , Filogenia , Sensibilidade e Especificidade , Esporotricose/microbiologia
19.
BMC Infect Dis ; 20(1): 126, 2020 Feb 11.
Artigo em Inglês | MEDLINE | ID: mdl-32046674

RESUMO

BACKGROUND: More and more azole-resistant strains emerged through the development of acquired resistance and an epidemiological shift towards inherently less susceptible species. The mechanisms of azoles resistance of Candida albicans is very complicated. In this study, we aim to investigate the mechanism of azole-resistant C. albicans isolated from the oral cavity of a patient with chronic mucocutaneous candidiasis (CMC). CASE PRESENTATION: CMC diagnosis was given based on clinical manifestations, laboratory test findings and gene sequencing technique. Minimum inhibitory concentration (MIC) of the fungal isolate, obtained from oral cavity termed as CA-R, was obtained by in vitro anti-fungal drugs susceptibility test. To further investigate the resistant mechanisms, we verified the mutations of drug target genes (i.e. ERG11 and ERG3) by Sanger sequencing, and verified the over-expression of ERG11 and drug efflux genes (i.e. CDR1 and CDR2) by RT-PCR. A heterozygous mutation of c.1162A > G resulting in p.K388E was detected in STAT1 of the patient. The expression of CDR1 and CDR2 in CA-R was 4.28-fold and 5.25-fold higher than that of type strain SC5314, respectively. CONCLUSIONS: Up-regulation of CDR1 and CDR2 was mainly responsible for the resistance of CA-R. For CMC or other immunodeficiency patients, drug resistance monitoring is necessary.


Assuntos
Antifúngicos/farmacologia , Azóis/farmacologia , Candida albicans/efeitos dos fármacos , Candidíase Mucocutânea Crônica/tratamento farmacológico , Candidíase Mucocutânea Crônica/microbiologia , Farmacorresistência Fúngica/genética , Mutação , Adolescente , Candida albicans/genética , Candida albicans/isolamento & purificação , Candidíase Mucocutânea Crônica/etiologia , Farmacorresistência Fúngica/efeitos dos fármacos , Proteínas Fúngicas/genética , Regulação Fúngica da Expressão Gênica/efeitos dos fármacos , Humanos , Masculino , Proteínas de Membrana Transportadoras/genética , Testes de Sensibilidade Microbiana , Boca/microbiologia
20.
Reprod Fertil Dev ; 32(18): 1350-1356, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33287951

RESUMO

Wild-type p53-induced phosphatase 1 (WIP1) plays an oncogenic function by increasing cell proliferation in various cancer types. Deficiency in WIP1 expression leads to male infertility, possibly by impairing the blood-testis barrier and spermatogenesis. However, how WIP1 functions in the Sertoli cells to affect male reproduction remains unclear. Thus, in the present study we used a swine Sertoli cell line to investigate whether WIP1 regulated the proliferation of Sertoli cells to participate in male reproduction. The WIP1 inhibitor GSK2830371, WIP1-short interference (si) RNAs and an upstream microRNA (miR-16) were used to inhibit the expression of WIP1, after which the proliferation of swine Sertoli cells, P53 expression and the levels of P53 phosphorylation were determined. Inhibiting WIP1 expression suppressed swine Sertoli cell proliferation, increased P53 expression and increased levels of P53 phosphorylation. In addition, overexpression of miR-16 in swine Sertoli cells resulted in a decrease in WIP1 expression and increases in both P53 expression and P53 phosphorylation. Together, these findings suggest that WIP1 positively regulates the proliferation of swine Sertoli cells by inhibiting P53 phosphorylation, and the miR-16 is likely also involved by targeting WIP1.


Assuntos
Proliferação de Células/genética , Proteína Fosfatase 2C/fisiologia , Células de Sertoli/fisiologia , Proteína Supressora de Tumor p53/fisiologia , Animais , Células Cultivadas , Regulação Enzimológica da Expressão Gênica , Masculino , MicroRNAs/fisiologia , Fosforilação , Proteína Fosfatase 2C/genética , Processamento de Proteína Pós-Traducional , Suínos , Proteína Supressora de Tumor p53/metabolismo
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