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1.
Biosens Bioelectron ; 150: 111862, 2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-31740256

RESUMO

Salmonella is the leading risk factor in food safety. Rapid, sensitive and accurate detection of Salmonella is a key to prevent and control the outbreaks of foodborne diseases caused by Salmonella. In this study, we reported a colorimetric biosensor for ultrasensitive detection of Salmonella Typhimurium using a magnetic grid separation column to efficiently separate target bacteria from large volume of sample and platinum loaded zeolitic imidazolate framework-8 (Pt@ZIF-8) nanocatalysts to effectively amplify biological signal. The target Salmonella cells in large volume of sample were first separated and concentrated using the magnetic grid separation column with immune magnetic particle chains, then conjugated with the immune Pt@ZIF-8 nanocatalysts to mimic peroxidase for catalysis of hydrogen peroxide-3,3',5,5'-tetramethylbenzidine, and finally determined by measuring the catalysate at characteristic wavelength of 450 nm. This proposed biosensor was able to separate ∼70% of target Salmonella cells from 50 mL of bacterial sample and quantitatively detect Salmonella from 101 to 104 CFU/mL in 2.5 h with the lower detection limit of 11 CFU/mL. The mean recovery for Salmonella in spiked chicken carcass was about 109.8%. This new magnetic grid separation method was first time reported for efficient separation of target bacteria from very large volume of sample to greatly improve the sensitivity of this biosensor and could be used with various biosensing assays for practical applications in routine detection of foodborne pathogens without any bacterial pre-enrichment.

2.
ACS Sens ; 5(1): 65-72, 2020 Jan 24.
Artigo em Inglês | MEDLINE | ID: mdl-31875386

RESUMO

Screening of pathogenic bacteria is a key to avoid food poisoning. The major drawbacks of existing assays for foodborne bacteria detection include long time for culture, complex DNA extraction for the polymerase chain reaction (PCR), and low sensitivity for enzyme-linked immunosorbent assay (ELISA), greatly limiting their practical applications. Here, we developed a sensitive optical biosensor based on porous gold@platinum nanocatalysts (Au@PtNCs) and a passive three-dimensional (3D) micromixer for fast detection of Salmonella typhimurium. The target Salmonella cells were first separated using immunomagnetic nanoparticles and the passive 3D micromixer. Then, immune Au@PtNCs were labeled onto the target cells as signal output to catalyze hydrogen peroxide-3,3',5,5'-tetramethylbenzidine. Finally, the absorbance was measured at 652 nm to calculate the bacterial amount. This optical biosensor could detect Salmonella at concentrations from 1.8 × 101 to 1.8 × 107 CFU/mL in 1 h. Its detection limit was calculated to be 17 CFU/mL. Besides, this passive 3D micromixer could magnetically separate 99% of target bacteria from the sample in 10 min. This biosensor has the potential to be extended to detect other bacteria by changing the antibodies.

3.
J Oral Rehabil ; 2019 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-31828830

RESUMO

BACKGROUND: Juvenile recurrent parotitis (JRP) is the second-most common childhood disease of the salivary glands after mumps. Since popularisation of mumps vaccination, children suffered from JRP more often, and the aetiology remains unclear. Chinese children had the habit of soft foods due to the special dietary habit of Asia. OBJECTIVES: To clarify whether mastication was related to the pathogenesis of JRP and whether the growth of salivary glands was influenced by soft diet. METHODS: Investigation of dietary habit and masticatory efficiency from 2015 to 2018 of children diagnosed with JRP compared with the normal children by the dentition. Mice had been fed a soft diet beginning in their development phase. The gland weight, amount of saliva, salivary amylase, histological and ultrastructural observation and the expression levels of EGF, FGFr2 and Wnt3a had been tested. RESULTS: The JRP children preferred soft foods and had a significantly lower masticatory efficiency than do normal children. When normalised by body weight, the gland weight, amount of saliva and amount of salivary amylase in the experimental group were significantly lower. The ultrastructural results showed that the acinar cells in the experimental groups were smaller and contained fewer electron-dense secretory granules than those in the control groups. The expression levels of EGF, FGFr2 and Wnt3a in the salivary glands of mice in the experimental groups were significantly lower than those of mice in the control groups. CONCLUSION: The soft diet indeed influenced the salivary gland through insufficient mastication, which could be one of the primary factors inducing JRP.

4.
J Autoimmun ; : 102358, 2019 Nov 19.
Artigo em Inglês | MEDLINE | ID: mdl-31757716

RESUMO

The hyperproliferation and hyperactivation of CD4+ T cells in salivary gland tissue is a hallmark of Sjögren's syndrome (SS). However, the role of long noncoding RNAs (lncRNAs) in the pathological process of SS and CD4+ T cell activation has not been fully elucidated. Here, we reported that lncRNA PVT1 was involved in the glycolytic metabolism reprogramming and proliferation upon CD4+ T cell activation. Expression of PVT1 was positively related with CD4+ T cell activation both in SS patients and Ex vivo antigen simulation. Depletion of PVT1 decreased the proliferation of murine CD4+ T cells and Jurkat T cells upon activation. We also showed that expression of the transcription factor Myc is regulated by PVT1 under antigen simulation. Depletion of PVT1 significantly decreased the expression of glycolytic genes, as well as several pivotal glycolytic proteins that were directly transcribed by Myc. Measurement of glucose content and lactate secretion indicated a defected lactate secretion and glucose uptake in PVT1-depleted T cells. Additionally, the real-time extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) measurement also affirmed that PVT1 maintains glycolytic levels, glycolytic capacity under stress and ECAR/OCR ratios during T cell activation. Polarizing assays indicate that PVT1 depletion defected the function of Th1 effector cells as well as down-regulated Myc expression and glycolytic levels. Furthermore, we observed increased glycolytic levels in CD4+ T cells from SS-like NOD/Ltj mice. Treatment with 2-deoxy-d-glucose (2-DG), an inhibitor of glycolysis, significantly decreased the extent of lymphocyte infiltration and CD4+ T cell numbers and attenuated the defect of salivary flow in the lesioned submandibular glands of NOD/Ltj mice. Thus, our study demonstrated that lncRNA PVT1, which was upregulated in the CD4+T cells of SS patients, could maintain the expression of Myc, thus controlling the proliferation and effector functions of CD4+ T cells through regulating the reprogramming of glycolysis. Inhibition of glycolysis could attenuate the proliferation of CD4+ T cells and the SS-like autoimmune response. Our study provides a novel mechanistic function of lncRNA PVT1 in the pathogenesis of SS.

5.
Mikrochim Acta ; 186(12): 757, 2019 11 09.
Artigo em Inglês | MEDLINE | ID: mdl-31707541

RESUMO

A disposable visual microfluidic immunosensor is described for the determination of foodborne pathogens using immunomagnetic separation, enzymatic catalysis and distance indication. Specifically, a sensor was designed to detect Salmonella typhimurium as a model pathogen. Magnetic nanoparticles (MNPs) were modified with the anti-Salmonella monoclonal antibodies and then used to enrich S. typhimurium from the sample. This is followed by conjugation to polystyrene microspheres modified with anti-Salmonella polyclonal antibodies and catalase to form the MNP-bacteria-polystyrene-catalase sandwich. The catalase on the complexes catalyzes the decomposition of hydrogen peroxide to produce oxygen after passing a micromixer. The generated oxygen gas increases the pressure in the chip and pushes the indicating red dye solution to travel along the channel towards the unsealed outlet. The travel distance of the red dye can be visually read and related to the amount of S. typhimurium using the calibration scale. The sensor can detect as low as 150 CFU·mL-1 within 2 h. Graphical abstractSchematic representation of the distance-based microfluidic immunosensor for visual detection of foodborne bacteria using immunomagnetic nanoparticles for bacteria separation, catalase for decomposition of hydrogen peroxide to form oxygen which causes a pressure increase, and red dyed particles movement for distance indication.

6.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 36(11): 1097-1099, 2019 Nov 10.
Artigo em Chinês | MEDLINE | ID: mdl-31703134

RESUMO

OBJECTIVE: To explore the genetic etiology of two unrelated patients with dyschromatosis symmetrica hereditaria. METHODS: Variant analysis of the ADAR gene was carried out by Sanger sequencing. RESULTS: Patient 1 was found to harbor a c.2633_2634delCT (p.Ser878fs) in exon 8 of the ADAR gene. The same variant was not found among 100 unrelated individuals. No pathogenic variant of the ADAR gene was found in patient 2. Functional prediction of the ADAR c.2633_2634delCT (p.Ser878fs) variant indicated it to be pathogenic by losing a catalytic structural domain. CONCLUSION: The c.2633_2634delCT (p.Ser878fs) variant of the ADAR gene probably underlies the pathogenesis of DSH in one of the patients.


Assuntos
Adenosina Desaminase/genética , Transtornos da Pigmentação/congênito , Proteínas de Ligação a RNA/genética , Humanos , Mutação , Linhagem , Transtornos da Pigmentação/genética , Tomografia Computadorizada por Raios X
7.
Genomics ; 2019 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-31626898

RESUMO

The egg parasitoid, Trichogramma chilonis, has significant control effects on agriculture and forestry pests and is widely employed in southern China for the biological control of lepidopteran pests. In this study, transcriptomic analysis was used to gain a clear understanding of the molecular changes in prepupae and pupae of T. chilonis. A total of 16.88 Gb of clean data were obtained and finally assembled into 43,136 unigenes, 18,880 of which were annotated. After FPKM standardization, 117 and 838 specific expression genes were found in prepupae and pupae, respectively. There were 3129 differentially expressed genes between prepupae and pupae. Compared to pupae, 806 genes were up-regulated and 2323 were down-regulated in prepupae. Background on the T. chilonis transcriptome, the enriched GO function and KEGG pathway analysis of DEGs were considered. As indicated by GO classification, up-regulated genes were mainly involved in chitin metabolism, cell adhesion and endocytic, while most down-regulated genes were involved in synthesis of cell components, ion transport and biological regulation. KEGG enrichment analysis showed that 458 DEGs were enriched in 94 metabolic pathways. DEGs involved in nucleotide replication and transcription, substance metabolism, insect hormone biosynthesis, cell growth and death, reproductive metabolism, circadian rhythms and signal transduction pathways were up-regulated or down-regulated to different degrees, indicating that these genes played important roles during the process of metamorphosis in T. chilonis. This study provides a rich data source for the future study of T. chilonis molecular and biological mechanisms. A large number of genes related to metamorphosis were found based on comparison analysis between prepupae and pupae transcriptomes. This study lays a good foundation for in-depth study of gene transcription and regulation mechanisms during T. chilonis metamorphosis.

8.
Fertil Steril ; 112(5): 882-891.e1, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31551156

RESUMO

OBJECTIVE: To investigate the possible impact of local inflammation on granulosa cells (GCs) and follicular development in endometriosis patients. DESIGN: Prospective study with related paired design. SETTING: Reproductive medicine center. PATIENT(S): A total of 80 endometriosis patients and 104 controls, with cultured GCs collected from control participants younger than 35 years. INTERVENTION(S): Tumor necrosis factor-α (TNF-α) and nuclear factor κB (NF-κB) inhibitor. MAIN OUTCOME MEASURE(S): Intrafollicular concentrations of cytokines measured with ELISA, NF-κB binding levels with electrophoretic mobility shift assay (EMSA), and telomerase activity (TA) with quantitative-telomeric repeat amplification protocol (Q-TRAP) assay, and protein and mRNA expression with Western blot and polymerase chain reaction analyses, respectively. RESULT(S): Patients with endometriosis exhibited a statistically significantly lower antral follicle count (11.48 ± 8.11 vs. 15.68 ± 8.56), lower number of retrieved oocytes (8.28 ± 6.69 vs. 10.87 ± 6.26), and lower number of mature oocytes (6.67 ± 6.09 vs. 8.53 ± 5.69). The GCs from endometriosis patients showed higher NF-κB binding activity and increased expression of inhibitor of NF-κB kinase subunit ß (IKKß, 2.743-fold) and NF-κB inhibitor α (IκBα, 5.017-fold). Their NF-κB p65 expression was negatively associated with mature oocytes (bNF-κB' = -0.304, R2 = 0.195, R = 0.442) but positively associated with intrafollicular TNF-α (r = 0.37); TA showed a negative relationship with NF-κB binding levels (r = -0.667). Tumor necrosis factor-α induced expression of IκBα (5.408-fold) and NF-κB p65 (1.400-fold) but lowered human telomerase reverse transcriptase (hTERT) and TA levels (0.0009 vs. 0.5619) in cultured GCs. However, inhibiting NF-κB obviously increased hTERT expression (1.988-fold). CONCLUSION(S): Endometriosis showed activated NF-κB pathways in GCs, which might negatively affect TA and oocyte quality. Intrafollicular TNF-α might down-regulate TA and hTERT via NF-κB pathway, but further studies are required.

9.
Cell Biol Int ; 2019 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-31502734

RESUMO

The purpose of this study was to explore the potential function of interleukin-11 (IL-11) in the pathogenesis of primary Sjögren's syndrome (pSS) patients. Real-time polymerase chain reaction was performed to examine IL-11 expression in the labial glands of 30 pSS patients and 30 healthy controls. Immunohistochemistry was conducted to assess the distribution of IL-ll-positive cells in labial glands. The human salivary gland (HSG) cell line was used to study the effects of IL-11 on gland epithelial cells in vitro. Cell viability and cell proliferation were examined by CCK-8 kit and EdU assay, respectively. The population of apoptotic cells was detected in flow cytometry followed by Annexin V/PI and Hoechst staining. We found that the expression levels of IL-11 were remarkably decreased in pSS labial glands and were positively correlated with C-reactive protein levels and negatively correlated with rheumatoid factor levels. Fewer numbers of glandular epithelial cells were observed to be positively stained with IL-11 antibody in labial glands from pSS patients than those in healthy control patients. After IL-11 treatment, the viability and proliferation of HSG cells were significantly higher than those in the control group. The total apoptotic and necrotic rates of HSG cells in the group after IL-11 treatment were significantly lower. In conclusion, the results indicated that IL-11 promoted viability and proliferation and inhibited apoptotic and necrotic rates of glandular epithelial cells. In pSS, downregulated IL-11 might contribute to the apoptosis of salivary gland epithelial cells. However, it might be a potential target to alleviate the pathological atrophy of glandular epithelial cells in pSS patients.

10.
Hum Reprod ; 34(9): 1788-1798, 2019 Sep 29.
Artigo em Inglês | MEDLINE | ID: mdl-31407797

RESUMO

STUDY QUESTION: Is embryo vitrification associated with a higher risk of adverse perinatal outcomes than slow-freezing? SUMMARY ANSWER: Embryo vitrification was not associated with increased risks of adverse perinatal outcomes of pre-term birth (PTB), low birthweight (LBW), small for gestational age (SGA), large for gestational age (LGA) and macrosomia, as compared to slow-freezing. WHAT IS KNOWN ALREADY: Vitrification is becoming a widely adopted technology for embryo cryopreservation with higher embryo survival rate and live birth rate than the slow-freezing technique. However, limited data are currently available on risks of adverse perinatal outcomes following vitrification as compared to that of slow-freezing. The impact of vitrification on perinatal outcomes remains further to be elucidated. STUDY DESIGN, SIZE, DURATION: Six large reproductive medical centers in Guangdong province, Southeast of China, took part in this multicenter retrospective cohort study. Cohorts of 3199 live born singletons after Day 3 frozen-thawed embryo transfer (FET) cycles with either vitrification or slow-freezing between January 2011 and December 2015 were included in the study. Each patient only contributed one cycle per cohort and vanishing twins were excluded. Propensity score (PS) matching was used to control for potential confounding factors. PARTICIPANTS/MATERIALS, SETTING, METHODS: All live-born singletons following either a vitrified or a slow-frozen cleavage FET cycle during the period from 2011 to 2015 were analyzed. Perinatal outcomes of PTB, LBW, macrosomia, SGA and LGA were compared. The vitrified and slow-frozen cohorts were matched by propensity scores with a 1:1 ratio accounting for potential confounding factors associated with perinatal outcomes. These variables included baseline demographics (maternal age, BMI, education level, parity, type of infertility and cause of infertility), as well as IVF characteristics (insemination method, endometrial preparation protocol and embryo cryopreservation duration). MAIN RESULTS AND THE ROLE OF CHANCE: A total of 2858 cases from vitrified embryo transfer (ET) and 341 babies from the slow-freezing group were included. After PS matching, 297 pairs of newborns were generated for comparison. The median gestational age was 39 weeks for both cohorts and the birthweights were comparable (3187.7 ± 502.1 g in the vitrified group vs. 3224.6 ± 483.6 in the slow-freezing group, P>0.05). There were no significant differences between the two groups on the incidence of PTB (5.4% vs. 7.7%), LBW (6.7% vs. 5.7%), macrosomia (5.7% vs. 6.1%), SGA (12.5% vs. 8.4%) and LGA (6.4% vs. 8.1%). Parallel logistic regression analysis indicated that vitrification was non-inferior to slow-freezing method in terms of the occurrence of PTB (OR, 0.68 [95% CI, 0.35, 1.31]), LBW (OR, 1.19[0.61-2.32]), macrosomia (OR, 0.94 [0.48-1.86]), SGA (1.55[0.91-2.64]) and LGA (0.78[0.42-1.45]), P>0.05. Sex-stratified PS matching models with multivariable regression analysis further confirmed that vitrification did not increase the risks of above-mentioned adverse perinatal outcomes in either the male or female infant cohort. LIMITATIONS, REASONS FOR CAUTION: Although the analysis was adjusted for a number of important confounders, the hospital dataset did not contain other potential confounders such as the medical history and obstetrics outcomes of women during pregnancy to allow adjustment. In addition, the current findings are only applicable to cleavage stage FET, but not pronuclei stage or blastocyst stage ET. WIDER IMPLICATIONS OF THE FINDINGS: Vitrified ET, in comparison with slow-frozen ET, was not associated with increased risks of adverse neonatal outcomes. With its superiority on live birth rates and non-inferiority on safety perinatal outcomes, transition from slow-freezing to the use of vitrification for embryo cryopreservation is reassuring. Nonetheless, future research is needed for the long-term effects of vitrification method on offspring's health outcomes. STUDY FUNDING/COMPETING INTEREST(S): The study was funded by the National Key Research and Development Program (2016YFC100205), Guangzhou Science and Technology Project (201804020087), Guangdong Province Science and Technology Project (2016A020218008) and Guangdong Provincial Key Laboratory of Reproductive Medicine (2012A061400003). The authors have no conflicts of interest to declare.

11.
Biosens Bioelectron ; 140: 111333, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31153017

RESUMO

Early screening of foodborne pathogens is a key to ensure food safety. In this study, we developed a microfluidic biosensor for online and sensitive detection of Salmonella based on immunomagnetic separation, fluorescence labeling and smartphone video processing. First, the immune magnetic nanoparticles were used to specifically separate and efficiently concentrate the target bacteria and the magnetic bacteria were formed. Then, the magnetic bacteria were labeled with the immune fluorescent microspheres and the fluorescent bacteria were formed. Finally, the fluorescent bacteria were continuously injected into the microfluidic chip on the smartphone-based fluorescent microscopic system, and the fluorescent spots were online counted using the smartphone App based on inter-frame difference algorithm to obtain the amount of the target bacteria. Under the optimal conditions, this proposed biosensor was able to quantitatively detect Salmonella typhimurium ranging from 1.4 × 102 to 1.4 × 106 CFU/mL, and its lower detection limit was 58 CFU/mL. This biosensor could be extended for detection of multiple foodborne pathogens using different fluorescent materials.


Assuntos
Carga Bacteriana/instrumentação , Técnicas Biossensoriais/instrumentação , Dispositivos Lab-On-A-Chip , Salmonella typhimurium/isolamento & purificação , Smartphone/instrumentação , Desenho de Equipamento , Fluorescência , Contaminação de Alimentos/análise , Microbiologia de Alimentos , Inocuidade dos Alimentos , Humanos , Separação Imunomagnética , Limite de Detecção , Aplicativos Móveis , Infecções por Salmonella/microbiologia
12.
Toxins (Basel) ; 11(6)2019 06 05.
Artigo em Inglês | MEDLINE | ID: mdl-31195758

RESUMO

In this paper, a highly sensitive plasmonic enzyme-linked immunosorbent assay (pELISA) was developed for the naked-eye detection of fumonisin B1 (FB1). Glucose oxidase (GOx) was used as an alternative to horseradish peroxidase as the carrier of the competing antigen. GOx catalyzed the oxidation of glucose to produce hydrogen peroxide, which acted as a reducing agent to reduce Au3+ to Au on the surface of gold seeds (5 nm), This reaction led to a color change in the solution from colorless to purple, which was observable to the naked eye. Various parameters that could influence the detection performance of pELISA were investigated. The developed method exhibited a considerably high sensitivity for FB1 qualitative naked-eye detection, with a visible cut-off limit of 1.25 ng/mL. Moreover, the proposed pELISA showed a good linear range of 0.31-10 ng/mL with a half maximal inhibitory concentration (IC50) of 1.86 ng/mL, which was approximately 13-fold lower than that of a horseradish peroxidase- (HRP)-based conventional ELISA. Meanwhile, the proposed method was highly specific and accurate. In summary, the new pELISA exhibited acceptable accuracy and precision for sensitive naked-eye detection of FB1 in maize samples and can be applied for the detection of other chemical contaminants.

13.
J Gerontol Nurs ; 45(5): 39-45, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-31026331

RESUMO

The current study aimed to evaluate the multidimensional effects of an interdisciplinary care team in patients with Alzheimer's disease (AD). A total of 129 patients with AD were randomly assigned to an interdisciplinary care group (n = 69) or usual care group (n = 60). Behavioral and psychological symptoms of patients with AD were measured during a 6-month treatment period. No differences were found in the baseline characteristics between the interdisciplinary care and usual care groups. Compared to usual care, interdisciplinary care greatly increased patients' activities of daily living (ADL) scores when measured at 3 and 6 months (p < 0.001). Findings provide evidence that an interdisciplinary care team approach is beneficial in improving ADL performance; thus, an interdisciplinary care team should be implemented in the care arrangements for patients with AD. [Journal of Gerontological Nursing, 45(5), 39-45.].


Assuntos
Atividades Cotidianas , Doença de Alzheimer/enfermagem , Progressão da Doença , Enfermagem Geriátrica/organização & administração , Equipe de Assistência ao Paciente/organização & administração , Enfermagem Psiquiátrica/organização & administração , Resultado do Tratamento , Idoso , Idoso de 80 Anos ou mais , China , Feminino , Humanos , Estudos Interdisciplinares , Masculino
14.
J Assist Reprod Genet ; 36(4): 741-747, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30778819

RESUMO

PURPOSE: Empty follicle syndrome (EFS) is a complex reproductive disorder characterized by the repeated failure to aspirate oocytes from mature ovarian follicles during in vitro fertilization (IVF). In addition to some cases caused by iatrogenic problems and known genetic factors, there are still many unexplained aspects of EFS. Here, we aimed to assess the clinical and genetic characteristics of two EFS patients. METHODS: We have characterized two primary infertility patients with EFS in a nonconsanguineous family from China. Both the patients presented similar clinical phenotypes, that is a few granulosa cells but no oocytes could be retrieved during repeated cycles with normal follicular development, E2 levels, and bioavailable hCG plasma levels. Abnormal oocytes were obtained once or twice between multiple IVF cycles. We performed Sanger sequencing of the LHCGR and ZP1~ZP4 genes in the patients, and further bioinformatics analysis was performed to identify pathogenic elements in the genes. RESULTS: A novel mutation, c.181C>T (p.Arg61Cys), and a known mutation, c.1169_1176delTTTTCCCA (p.Ile390Thrfs*16), in the ZP1 gene were both identified in patient 2, but no mutations were identified in patient 1. The novel mutation inherited from her mother was absent in the control cohort and the ExAc database. The arginine residue is conserved at this position, and its replacement by cysteine was predicted to be deleterious. In another allele, a paternal frameshift mutation was predicted to introduce premature stop codons, resulting in the deletion of 234 amino acids from the C-terminus of the ZP1 protein. CONCLUSIONS: Our findings presented compound heterozygous mutations in ZP1 associated with EFS and abnormal oocytes and provided further new evidence for the genetic basis of EFS and support for the genetic diagnosis of infertile individuals.


Assuntos
Predisposição Genética para Doença , Infertilidade Feminina/genética , Doenças Ovarianas/genética , Glicoproteínas da Zona Pelúcida/genética , Adulto , China/epidemiologia , Feminino , Humanos , Infertilidade Feminina/patologia , Mutação , Oócitos/crescimento & desenvolvimento , Oócitos/patologia , Doenças Ovarianas/patologia , Folículo Ovariano/crescimento & desenvolvimento , Folículo Ovariano/patologia , Ovulação/genética , Indução da Ovulação/métodos , Fenótipo , Zona Pelúcida/patologia
15.
Exp Cell Res ; 375(2): 51-59, 2019 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-30610847

RESUMO

Abnormal signaling transduction in salivary gland cells is associated with the pathogenesis of Sjögren's syndrome (SS). Previously, we identified aberrant expression of toll-like receptor 9 (TLR9) in gland cells of SS patients and mouse models. In this study, we investigated the role of TLR9 and its downstream p38/mitogen-activated protein kinase (MAPK) and c-Jun N-terminal kinase (JNK) signaling in mediating apoptosis and autophagy in human salivary gland (HSG) cells. We selected either CpG-Odn, a classical TLR9 activator, or lentivirus-packaged TLR9 full-length cDNA to activate TLR9 signaling transduction. Activation of TLR9 signaling induced phosphorylation of its downstream protein kinases, p38/MAPK and JNK, in a time-dependent manner, and decreased HSG cell viability. Western blotting of LC3B-II and p62 in both normal and autophagic flux-administered conditions revealed elevated autophagy upon TLR9 activation. Observing the cell cytoplasm through transmission electron microscopy and mRFP-GFP-LC3B-tagged fluorescence confirmed an increased number of autophagosomes and autolysosomes in TLR9-activated cells. Bax/Bcl-2 ratio calculations, caspase-3 activity assays and Hoechst nuclear staining were utilized to confirm the involvement of apoptosis in TLR9 signaling activation. Furthermore, we selected SB239063, a p38/MAPK signaling inhibitor, and SP600125, a JNK inhibitor, to identify the functions of p38/MAPK and JNK in TLR9-mediated signaling transduction. Multiple approaches, including Western blotting assays, fluorescence assessments and caspase-3 activity measurements, confirmed that inhibition of p38/MAPK signaling ameliorated both autophagy and apoptosis in TLR9-activated HSG cells, whereas inhibition of JNK signaling attenuated apoptosis but failed to modulate autophagy in the models mentioned above. Our results indicate a divergent function of p38/MAPK and JNK in TLR9-mediated autophagy and apoptosis in salivary gland cells.

16.
Biosens Bioelectron ; 124-125: 143-149, 2019 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-30366259

RESUMO

We intended to develop a novel biosensor using gold nanoparticles (AuNPs) for indicating different concentrations of E. coli O157:H7 and smart phone imaging APP for monitoring color change of the AuNPs. The magnetic nanoparticles (MNPs) modified with the capture antibodies and the polystyrene microspheres (PSs) modified with the detection antibodies and the catalases were simultaneously used to react with the target bacteria in the first mixing channel of the microfluidic chip, and hydrogen peroxide was injected and catalyzed by the catalases on the MNP-bacteria-PS complexes. After the mixture of the AuNPs and the crosslinking agents were injected to react with the catalysate in the second mixing channel and incubated in the detection chamber, the aggregation of the AuNPs was triggered through the crosslinking agents, resulting in the color of the AuNPs changing from blue to red. Finally, the color was measured using the smart phone imaging APP to determine the amount of the bacteria. This biosensor exhibited a good specificity and sensitivity for detection of E. coli O157:H7 in chicken samples with a lower detection limit of 50 CFU/mL.


Assuntos
Técnicas Biossensoriais , Infecções por Escherichia coli/diagnóstico , Escherichia coli O157/isolamento & purificação , Microbiologia de Alimentos , Anticorpos Imobilizados/química , Colorimetria , Infecções por Escherichia coli/microbiologia , Escherichia coli O157/patogenicidade , Ouro/química , Humanos , Nanopartículas Metálicas/química , Microfluídica/instrumentação , Smartphone
17.
Micromachines (Basel) ; 9(12)2018 Nov 27.
Artigo em Inglês | MEDLINE | ID: mdl-30486364

RESUMO

Immunomagnetic separation has been widely used for the separation and concentration of foodborne pathogens from complex food samples, however it can only handle a small volume of samples. In this paper, we presented a novel fluidic device for the specific and efficient separation and concentration of salmonella typhimurium using self-assembled magnetic nanoparticle chains. The laminated sawtooth-shaped iron foils were first mounted in the 3D-printed matrix and magnetized by a strong magnet to generate dot-array high gradient magnetic fields in the fluidic channel, which was simulated using COMSOL (5.3a, Burlington, MA, USA). Then, magnetic nanoparticles with a diameter of 150 nm, which were modified with the anti-salmonella polyclonal antibodies, were injected into the channel, and the magnetic nanoparticle chains were vertically formed at the dots and verified using a fluorescence inverted microscope. Finally, the bacterial sample was continuous-flow injected, and the target bacteria could be captured by the antibodies on the chains, followed by gold standard culture plating to determine the amount of the target bacteria. Under the optimal conditions, the target bacteria could be separated with a separation efficiency of 80% in 45 min. This fluidic device could be further improved using thinner sawtooth-shaped iron foils and stronger magnets to obtain a better dot-array magnetic field with larger magnetic intensity and denser dot distribution, and has the potential to be integrated with the existing biological assays for rapid and sensitive detection of foodborne bacteria.

18.
J Craniomaxillofac Surg ; 46(11): 1899-1904, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30249485

RESUMO

OBJECTIVE: The objective of this study was to analyze bone marrow stromal antigen-2 (BST-2) levels in labial glands, total peripheral blood mononuclear cells (PBMCs) and PBMC subpopulations from primary Sjögren's syndrome (pSS) patients and determine the correlation between BST-2 expression and clinical characteristics. MATERIALS AND METHODS: PBMC subsets were positively separated using magnetic microbeads. BST-2 mRNA levels in labial glands, total PBMCs and PBMC subsets of 30 pSS and 30 healthy control (HC) subjects were investigated using real-time polymerase chain reaction (RT-PCR). Distribution of BST-2-positive cells in the labial glands was assessed by immunohistochemistry. RESULTS: BST-2 was significantly increased in pSS labial glands and was positively correlated with the VAS value for parotid gland swelling and rheumatoid factor and ß2-microglobulin serum levels. BST-2 levels were statistically different between pSS patients with positive and negative expression of anti-SSA antibody. Positive focal infiltrating lymphocytes and adjacent ductal epithelial cells were observed in labial glands from pSS patients, while there were a few scattered positive ductal epithelial cells in controls. BST-2 was also up-regulated in CD19+ B cells and the remaining CD4-CD8-CD19- PBMCs. CONCLUSION: BST-2 was aberrantly expressed in pSS patients, and expression in labial glands was positively correlated with important clinical characteristics; thus, it may be a potential biomarker of pSS activity.


Assuntos
Antígenos CD/metabolismo , Síndrome de Sjogren/metabolismo , Adulto , Idoso , Estudos de Casos e Controles , Feminino , Proteínas Ligadas por GPI/metabolismo , Humanos , Contagem de Leucócitos , Leucócitos Mononucleares , Lábio/metabolismo , Pessoa de Meia-Idade , Síndrome de Sjogren/imunologia , Síndrome de Sjogren/patologia , Adulto Jovem
19.
J Assist Reprod Genet ; 35(8): 1537-1542, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29926375

RESUMO

PURPOSE: The objective of this study was to determine whether ammonium accumulates in IVF media during fertility process and whether the brief co-incubation of gametes (bIVF) benefited the outcomes of newborns. METHODS: Ammonium levels in IVF media during gamete co-incubation were measured and the effects of bIVF on neonatal outcomes were evaluated retrospectively in this study. RESULTS: A total of 609 live newborns cycles were included in this study. The results showed that ammonium levels in the conventional IVF (cIVF) media was significantly increased than that in bIVF and control media (27.32 ± 5.60 vs 20.71 ± 3.89, P = 0.03; 27.32 ± 5.60 vs 19.46 ± 1.31, P = 0.01, respectively). In the cIVF group, the mean gestational age was significantly lower (37.36 ± 2.29 vs. 37.74 ± 1.94 weeks, P = 0.031) and the incidence of preterm birth (< 37 weeks) was higher than that in the bIVF group (25.80 vs. 17.63%, P = 0.015). Singleton cycles and twin cycles were then analyzed respectively. The gestational age and birth weight of the singleton cycles were similar between the two groups. However, of the twin cycles, the gestational age was significantly decreased and the rate of preterm birth was increased significantly in the cIVF group (35.76 ± 2.31 vs. 36.48 ± 1.73, P = 0.013; 53.33 vs. 31.52%, P = 0.002, respectively). CONCLUSIONS: There is an ammonium accumulation in IVF media during co-incubation of gametes. And bIVF reduces the risk of preterm birth (< 37 weeks), especially with regard to preterm birth of the twin cycles, and seems to be a safe alternative method for improving the neonatal outcomes compared with cIVF.


Assuntos
Transferência Embrionária/métodos , Fertilização In Vitro , Células Germinativas/citologia , Adulto , Peso ao Nascer , Feminino , Idade Gestacional , Humanos , Recém-Nascido , Nascimento Vivo , Gravidez , Taxa de Gravidez , Nascimento Prematuro , Gêmeos
20.
J Assist Reprod Genet ; 35(7): 1349-1356, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29704226

RESUMO

PURPOSE: Cleavage of the zygote during human reproduction is a key event of early embryonic development. The genetic events associated with idiopathic embryonic cleavage failure are not certain. Mutations in the tubulin beta 8 class VIII (TUBB8) gene have been reported to be associated with oocyte maturation, fertilization, and developmental arrest. Here, we aimed to assess the clinical and genetic characteristics of complete cleavage failure in fertilized eggs. METHODS: We have characterized a patient with a 9-year history of primary infertility in a non-consanguineous family from China. The patient presented complete cleavage failure in all two-pronuclear (2PN) fertilized oocytes after 2 cycles of in vitro fertilization (IVF). We performed Sanger sequencing of the TUBB8 gene in the patient, and further bioinformatics analysis to identify pathogenesis of gene. RESULTS: A novel homozygous mutation, c.322G > A (p.Glu108Lys), was detected, and this change was absent from 179 control subjects. Glutamic acid is highly conserved at this position, and replacement by lysine was predicted to be repelled by the α-tubulin positive region, disrupting the α-ß tubulin interaction. CONCLUSIONS: Our findings presented a homozygous mutation of TUBB8 associated with complete cleavage failure in fertilized eggs and provided new data for the genotype-phenotype of TUBB8-related diseases.


Assuntos
Fase de Clivagem do Zigoto/fisiologia , Desenvolvimento Embrionário/fisiologia , Mutação/genética , Oócitos/fisiologia , Tubulina (Proteína)/genética , Adulto , China , Feminino , Fertilização In Vitro/métodos , Humanos , Infertilidade Feminina/genética , Zigoto/fisiologia
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