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1.
Clin Chim Acta ; 501: 12-19, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31805271

RESUMO

BACKGROUND: We investigated the characteristics of antineutrophil cytoplasmic antibody (ANCA)-associated glomerulonephritis (ANCA-GN) and immunoglobulin G4 (IgG4)-related kidney disease (IgG4-RKD) overlap syndrome. METHODS: This is a 2-center study with 19 patients. RESULTS: Fifteen patients were classified as microscopic polyangiitis (MPA). The initial serum creatinine levels were 320.9 ± 191.4 µmol/l and the BVAS was 19.7 ± 7.4 at diagnosis. Hematuria was absent or slight in 13 (68.4%) cases. Renal histology of these patients revealed concurrent ANCA-GN and IgG4-RKD. Regarding the histological classification of ANCA-GN, 9 (47.4%), 8 (42.1%), 1 (5.3%) and 1 (5.3%) patients were classified as focal, crescentic, mixed and sclerotic ANCA-GN, respectively. MPO-ANCA could be detected in 17/19 (89.5%) patients. IgG subclasses of MPO-ANCA were tested in 10 patients, and all were positive for IgG4-MPO-ANCA. In patients with combined ANCA-GN and IgG4-RKD, the percentage of positive IgG1-MPO-ANCA was significantly lower than the control group of 20 AAV patients without IgG4-RKD (P = 0.002), and the percentage of positive IgG4-MPO-ANCA was higher than the control group, but this difference was not statistically significant (P = 0.14). CONCLUSIONS: ANCA-GN and IgG4-RKD overlap syndrome concerned mainly MPO-ANCA positive patients. The IgG subclass analysis of MPO-ANCA showed lower percentage of IgG1 subclass. The association between ANCA-GN and IgG4-RKD is possible and represents a special entity.

2.
Artif Cells Nanomed Biotechnol ; 48(1): 276-287, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31858826

RESUMO

Background: The abnormal expression Dopamine D1 receptor (DRD1) gives rise to the dysfunction of dopaminergic neurotransmitter and may be associated with the occurrence of schizophrenia. MicroRNAs (miRNAs) can regulate the DRD1 expression by binding 3'UTR and be involved in the post-transcriptional regulation.Methods: We first constructed the pmirGLO-recombined vectors of series of DRD1 gene 3'UTR-truncated fragments and performed the luciferase receptor assay to screen the underlying 3'UTR sequence targeted by miRNAs. Then, we predicted the potential miRNAs binding the target sequence and confirmed their effects using luciferase receptor assay after transfection of the miRNA mimics/inhibitors. We also examined the effects of the miRNA on the endogenous DRD1 expression.Results: We found that the DRD1 3'UTR ranging from -12 to +1135 bp was essential for the post-transcriptional regulation of miRNAs. The deletion of -12 to +154 bp fragment significantly increased the luciferase expression but not the mRNA expression. The miRNA-15a, miRNA-15b and miRNA 16 affected DRD1 expression in HEK293, U87, SK-N-SH and SH-SY5Y cell lines.Conclusion: The miRNA-15a, miRNA-15b and miRNA-16 inhibit the human dopamine D1 receptor expression by targeting 3'UTR -12 to +154 bp.HighlightsDRD1 3'UTR ranging from -12 to +1135 bp was essential for the post-transcriptional regulation of miRNAs.The deletion of -12 to +154 bp fragment significantly increased the luciferase expression but not the mRNA expression.The miRNA-15a, miRNA-15b and miRNA 16 affected DRD1 expression in different cell lines, respectively.

3.
J Orthop Surg Res ; 14(1): 416, 2019 Dec 09.
Artigo em Inglês | MEDLINE | ID: mdl-31815638

RESUMO

BACKGROUND: Long non-coding RNA (LncRNA) TUG1 plays critical roles in the development of human cancers. Its inhibition has been proved to participate in ankylosing spondylitis, which is an inverse pathological procedure of osteoporosis. In the present study, we aim to investigate the role of lncRNA TUG1 in ankylosing spondylitis. MATERIALS AND METHODS: Expressions of lncRNA TUG1 in plasma of 98 patients with osteoporosis and 60 healthy participants were detected by real-time quantitative PCR (RT-qPCR). Diagnostic values of lncRNA CASC11 for osteoclasts were performed by the ROC curve with osteoporosis patients as positive and healthy participants as negative. All experiments were repeated 3 times. Mean ± standard deviation was calculated. RESULTS: We found that plasma lncRNA TUG1 was upregulated in osteoporosis patients than in healthy participants. Upregulation of plasma lncRNA TUG1 distinguished osteoporosis patients from healthy participants. LncRNA TUG1 level increased with the advances of clinical stages. Over-expression of lncRNA TUG1 promoted the proliferation and inhibited the apoptosis of mice osteoclasts, while lncRNA TUG1 siRNA silencing played an opposite role. In addition, lncRNA TUG1 over-expression led to downregulated PTEN, while lncRNA TUG1 siRNA silencing played an opposite role. CONCLUSION: Therefore, lncRNA TUG1 is upregulated in osteoporosis and regulates the proliferation and apoptosis of osteoclasts. lncRNA TUG1 knockdown may serve as a promising therapeutic target for osteoporosis by inhibiting the proliferation and promoting the apoptosis of osteoclasts through PTEN.

4.
Medicine (Baltimore) ; 98(47): e17901, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31764787

RESUMO

BACKGROUND: Salpingectomy is routinely performed in ectopic pregnancy (EP). However, the effect of the surgery on the ovarian reserve and ovarian response in EP patients is still uncertain and has not been systematically evaluated. Therefore, we conducted this meta-analysis to provide a comparison of the ovarian reserve and ovarian response between the pre-salpingectomy and post-salpingectomy in EP patients. METHODS: Pubmed, Embase, and Cochrane Library were searched for all relevant articles published up to December 2018. We retrieved the basic information and data of the included studies. The data was analyzed by Review Manager 5.3 software (Cochrane Collaboration, Oxford, UK). RESULTS: A total of 243 articles were extracted from the databases, and 7 studies were included in the meta-analysis. The ovarian reserve including anti-Mullerian hormone (inverse variance [IV] -0.7 [95% confidence interval [CI] -0.63, 0.49]), antral follicle count (IV 1.7 [95% CI -2.02, 5.42]) and basal follicle stimulating hormone (IV 0.02 [95% CI -0.63, 0.68]) was comparable between the pre-salpingectomy group and the post-salpingectomy group. The amount of gonadotropin was significantly higher in the post-salpingectomy group when compared with that in the pre-salpingectomy group (IV -212.65 [95% CI -383.59, -41.71]). There was no significant difference in the left parameters of the ovarian response including the duration of gonadotropin stimulation (IV -0.32 [95% CI -0.76, 0.12]), the estrogen level on the human chorionic gonadotropin triggering day (IV -4.12 [95% CI -236.27, -228.04]) and the number of retrieved oocytes (IV 0.35 [95% CI -0.76, 1.46]) between 2 groups. CONCLUSIONS: The current results suggest that salpingectomy has no negative effect on the ovarian reserve and ovarian response.


Assuntos
Reserva Ovariana , Ovário/fisiopatologia , Gravidez Ectópica/fisiopatologia , Gravidez Ectópica/cirurgia , Salpingectomia , Feminino , Humanos , Gravidez
5.
Proc Natl Acad Sci U S A ; 116(47): 23467-23472, 2019 Nov 19.
Artigo em Inglês | MEDLINE | ID: mdl-31690659

RESUMO

We present a theory on the coalescence of 2 spherical liquid droplets that are initially stationary. The evolution of the radius of a liquid neck formed upon coalescence was formulated as an initial value problem and then solved to yield an exact solution without free parameters, with its 2 asymptotic approximations reproducing the well-known scaling relations in the inertially limited viscous and inertial regimes. The viscous-to-inertial crossover observed in previous research is also recovered by the theory, rendering the collapse of data of different viscosities onto a single curve.

6.
Neurosci Lett ; 713: 134535, 2019 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-31586698

RESUMO

The Schizophrenia Psychiatric GWAS Consortium (PGC) has identified the rs1625579 polymorphism in the MIR137 gene, which encodes miR-137, as the strongest new association with schizophrenia in the European population. However, whether the influence of rs1625579 on schizophrenia in the Asian population is consistent with these results remains unclear. A total of 21 studies (9878 schizophrenic patients and 9447 control subjects) that met the inclusion criteria were included in our meta-analysis. Pooled analysis, subgroup analysis, sensitivity analysis and publication bias were performed. The T allele, TT genotype and TT + GG genotype were associated with schizophrenia as risk factors. Subgroup analysis shows that no heterogeneity existed in the European and Asian populations. Our meta-analysis found that the Rs1625579 polymorphism in the MIR137 gene was associated with the risk of schizophrenia. The current findings provide a reference for case-control studies of schizophrenia in the future.

7.
J Mol Neurosci ; 2019 Aug 22.
Artigo em Inglês | MEDLINE | ID: mdl-31440993

RESUMO

The dopamine transporter is coded by the SLC6A3 gene and plays an important role in regulation of the neurotransmitter dopamine. To detect the association between the SLC6A3 gene and the risk of schizophrenia, 31 case-control articles were included in this meta-analysis. There were 23 studies with 40 bp VNTR (3246 cases and 3639 controls), 4 studies with rs40184 (2020 cases and 1674 controls), rs6347 (1317 cases and 1917 controls), rs403636 (2045 cases and 1704 controls), and rs2975226 (849 cases and 904 controls); and 3 studies with rs12516948 (1920 cases and 1569 controls), rs27072 (984 cases and 1015 controls), rs6869645 (1142 cases and 1082 controls), rs37022 (1168 cases and 1091 controls), rs464049 (1169cases and 1096 controls), rs2652511 (707 cases and 714 controls), and rs3756450 (1176 cases and 1096 controls). Pooled, subgroup, and sensitivity analyses were performed, and the results were visualized by forest and funnel plots. In the dominant genetic model, the genotype AA+AT of rs2975226 in the Indian population (Pz = 0, odds ratio [OR] = 3.245, 95% confidence interval [CI] = 1.806-5.831), TT of rs464049 (Pz = 0.002, OR = 1.389, 95% CI = 1.129-1.708), and TT of rs3756450 (Pz = 0.014, OR = 1.251, 95% CI = 1.047-1.495) might be risk factors for schizophrenia. Additionally, no other single nucleotide polymorphisms were observed. These results indicate that more functional studies are warranted.

8.
J Chin Med Assoc ; 82(11): 845-848, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31453864

RESUMO

BACKGROUND: Ovarian stimulation with clomiphene (CC) or progestin has been applied for patients with diminished ovarian reserve (DOR). However, it remains unclear which treatment confers greater benefits. This study aimed to compare the outcomes of progestin-primed ovarian stimulation (PPOS) protocol vs CC-primed ovarian stimulation (CPOS) in infertile women with DOR. METHODS: A before-and-after self-controlled study was conducted to retrospectively investigate the data from 50 infertile women with DOR, who failed to conceive in their first in vitro fertilization/intracytoplasmic sperm injection-frozen embryo transfer cycle when stimulated with CPOS, and switched to PPOS, in the Reproductive Medicine Center of Changzhou Maternal and Child Health Care Hospital. RESULTS: Our results showed that PPOS significantly suppressed the luteinizing hormone (LH) surge and yielded more satisfactory results in patients with DOR, including increased number of retrieved oocytes, MII mature oocytes, normal fertilized oocytes, cleaved embryos, high-grade embryos, cryopreserved embryos, pregnancy rate, live-birth rate, and decreased miscarriage rates. CONCLUSION: Our study demonstrated that compared with CPOS protocol, PPOS protocol could not only suppress the LH surge but also improved the quantity, particularly the quality of oocytes in patients with DOR, suggesting that PPOS treatment is more effective than CPOS for patients with DOR.

10.
Mol Med Rep ; 20(2): 1187-1195, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31173235

RESUMO

The aim of the present study was to investigate the effects of alisol B 23­acetate (AB23A) on inhibiting the viability and inducing apoptosis of human non­small cell lung cancer (NSCLC) cells and the anticancer mechanisms of AB23A in vitro. The viability of A549 cells following treatment with different doses of AB23A was examined using a Cell Counting Kit­8 assay. Subsequently, apoptosis and the cell cycle were detected using flow cytometric analysis. The effect of AB23A on migration and invasion of A549 cells was detected by wound healing and Transwell assays. Western blotting was performed to determine the relative expression of Bax/Bcl­2, phosphatidylinositol 3­kinase (PI3K), protein kinase B (AKT) and mammalian target of rapamycin (mTOR). AB23A markedly inhibited the viability enhanced apoptosis of A549 cells and arrested the cell cycle in G1 phase. Additionally, AB23A upregulated the ratio of Bax/Bcl­2 in the A549 cells in a concentration­dependent manner. The results of wound healing and Transwell assays indicated that AB23A also suppresses the migration and invasion ability of A549 cells. Furthermore, AB23A reduced the protein levels of phosphorylated AKT, PI3K and mTOR. In conclusion, AB23A exerted anti­cancer activity via inhibiting cells viability, migration and invasion, and promoting apoptosis. Therefore, AB23A is a potential antitumor drug for the treatment of NSCLC.


Assuntos
Apoptose , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Colestenonas/farmacologia , Neoplasias Pulmonares/tratamento farmacológico , Transdução de Sinais , Células A549 , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Carcinoma Pulmonar de Células não Pequenas/fisiopatologia , Linhagem Celular , Movimento Celular , Sobrevivência Celular , Colestenonas/uso terapêutico , Humanos , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/fisiopatologia , Invasividade Neoplásica , Fosfatidilinositol 3-Quinase/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Serina-Treonina Quinases TOR/metabolismo
11.
Artigo em Inglês | MEDLINE | ID: mdl-31233944

RESUMO

An ultra-high-performance liquid chromatography-tandem mass spectrometry method was developed to analyze cephalexin in swine tissues, urine, and feces. Samples were extracted with 1% sulfuric acid, followed by purification using MCX cartridges. Mean recoveries were 95.4%-100.7% with inter-day relative standard deviations of <8.6%. The quantitation limit was 5 µg/kg for fat and urine, and 10 µg/kg for muscle, liver, kidney, and feces. Cephalexin residue depletion was determined using 32 healthy pigs, randomly divided into eight (seven treated and one control) groups. Treated groups were intramuscularly administered 10 mg/kg b.w. five times at 24-h intervals and euthanized 6 h and 1, 2, 3, 5, 7, and 10 days after the last injection. Cephalexin was eliminated rapidly in swine muscle, liver, fat, and feces. The highest concentrations among edible organs were detected in the kidney. Moreover, the longest elimination period of cephalexin in swine was determined in urine. These results indicated that kidney and urine were likely target matrices for cephalexin residue detection in swine.


Assuntos
Antibacterianos/análise , Cefalexina/análise , Cromatografia Líquida de Alta Pressão/métodos , Resíduos de Drogas/análise , Espectrometria de Massas em Tandem/métodos , Animais , Antibacterianos/urina , Cefalexina/urina , Gorduras/química , Fezes/química , Rim/química , Fígado/química , Músculos/química , Suínos
12.
Medicine (Baltimore) ; 98(19): e15512, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31083194

RESUMO

At present, the precise role of human menopausal gonadotropin (HMG) and recombinant luteinizing hormone (rLH) supplementation at an early time of follicular phase on in vitro fertilization (IVF)/intra cytoplasmatic sperm injection (ICSI) outcomes remains uncertain.Here infertile women of normal ovarian function undergoing their first cycle of IVF/ICSI were studied and were randomly allocated into 3 groups. Group 1, ovarian stimulation with 150IU recombinant follicle-stimulating hormone (FSH) alone. Group 2, patients received 75IU rFSH and 75IU HMG. Group 3 patients were given 150IU rFSH and 75IU rLH.There were no significant differences in the clinical characteristics, ovarian response, the biochemical, clinical and ongoing pregnancy rates among the 3 groups. No significant differences were found in biochemical, clinical and ongoing pregnancy rates between the patients whose LH levels were lower than 0.75 mIU/ml and those above this threshold in group 1. Furthermore, there were also no significant differences in biochemical, clinical and ongoing pregnancy rates among the 3 group patients whose LH level lower than 0.75 mIU/ml.The results showed that either the addition of HMG or rLH supplementation at an early time of follicular phase produce no significant benefit on IVF outcome in patients with normal ovarian function.


Assuntos
Fármacos para a Fertilidade Feminina/administração & dosagem , Hormônio Luteinizante/administração & dosagem , Menotropinas/administração & dosagem , Indução da Ovulação/métodos , Adulto , Feminino , Fertilização In Vitro , Fase Folicular , Humanos , Gravidez , Taxa de Gravidez , Estudos Retrospectivos , Adulto Jovem
13.
Environ Int ; 127: 361-370, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30954722

RESUMO

BACKGROUND: The extensive use of colistin in swine production may have contributed to the recent emergence of corresponding mobile resistance gene mcr-1. The use of colistin as a feed additive was banned in China in April 2017. OBJECTIVES: To examine the occurrence of colistin and dissemination of mcr-1 in swine feedlots before and after the colistin ban and effects of different manure treatments. METHODS: Environmental samples were collected from swine feedlots before (December 2016) and after (December 2017) the colistin ban. Colistin concentrations were determined by ultra-high performance liquid chromatography coupled to tandem mass spectrometry. The prevalence of mcr-1 were determined by quantitative PCR analysis, while bacterial community composition was investigated by 16S rRNA sequencing. RESULTS: In 2016, colistin was detected in feed and fresh manure samples at 67 mg/kg and 17 mg/kg, respectively, but was absent from all samples in 2017. In 2016, the relative abundance of mcr-1 in fresh manure was lower than that in solid samples after natural drying, while a higher relative abundance was detected in fresh manure samples compared with biogas slurry samples. A strong correlation between colistin concentration and relative abundance of mcr-1 was observed in fresh manure. The samples collected in 2017 showed a lower relative abundance of mcr-1 compared with those collected in 2016. Bacterial community analysis showed that the abundance of Enterobacteriaceae, which act as a vehicle and reservoir of mcr-1, increased with natural dying but decreased with anaerobic digestion. CONCLUSIONS: The presence of colistin exerts direct selection pressure for the accumulation of mcr-1 in manure, while the ban on colistin likely halted the dissemination of mcr-1 on pig farms. Anaerobic digestion is an effective waste treatment process for removing mcr-1, which might be mainly driven by the shift in bacterial community structure.


Assuntos
Bactérias/efeitos dos fármacos , Colistina/química , Esterco/análise , Esterco/microbiologia , Suínos , Animais , Antibacterianos/análise , Antibacterianos/química , Antibacterianos/farmacologia , China , Colistina/análise , Farmacorresistência Bacteriana/efeitos dos fármacos , RNA Ribossômico 16S , Suínos/metabolismo , Suínos/microbiologia
14.
Mol Genet Genomic Med ; 7(5): e652, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30908890

RESUMO

BACKGROUND: This study investigated the effects of haplotypes T-G and C-A derived from NG_012836.1:g.4160T>C and NG_012836.1:g.4326G>A on protein expression levels in vitro and identified the functional sequence in the regulatory region of the GABRB3 gene linked to possible associations with schizophrenia. METHODS: Recombinant plasmids with haplotypes T-G and C-A and 10 recombinant vectors containing deletion fragments from the GABRB3 gene 5' regulatory region were transfected into HEK-293, SK-N-SH, and SH-SY5Y cells. The relative fluorescence intensity of the two haplotypes and different sequences was compared using a dual luciferase reporter assay system. RESULTS: The relative fluorescence intensity of haplotype C-A was significantly lower than that of T-G. We shortened the core promoter sequence of the GABRB3 gene 5' regulation region from -177 bp to -18 bp (ATG+1). We also found an expression suppression region from -1,735 bp to -1,638 bp and an enhanced regulatory region from -1,638 bp to -1,335 bp. Multiple inhibitory functional elements were identified in the region from -680 bp to -177 bp. CONCLUSION: We demonstrated that haplotype C-A might increase the risk of schizophrenia and found multiple regulatory regions that had an effect on GABRB3 receptor expression.


Assuntos
Receptores de GABA-A/genética , Esquizofrenia/genética , Região 5'-Flanqueadora , Linhagem Celular Tumoral , Células HEK293 , Haplótipos , Humanos , Regiões Promotoras Genéticas , Receptores de GABA-A/metabolismo
15.
Mol Cell Endocrinol ; 490: 15-20, 2019 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-30922932

RESUMO

The steroidogenic acute regulatory protein (STAR) governs the rate-limiting step in steroidogenesis, and its expression varies depending on the needs of the specific tissue. It is well known that tight control of steroid production is essential for multiple processes involved in reproduction. We recently showed that Star is regulated at the posttranscriptional level in vitro by H19 and let-7. Here we demonstrate that this novel regulatory mechanism is functional in vivo, regulated by cAMP, and that loss of H19 not only disrupts ovarian STAR but also results in altered progesterone production in an H19KO mouse model. This work further strengthens the possibility that noncoding-RNA-mediated regulation of STAR may play an important role in the regulation of steroid hormone production, and contributes further to our understanding of the many ways in which this important gene is regulated.


Assuntos
Ovário/metabolismo , Fosfoproteínas/metabolismo , Progesterona/biossíntese , RNA Longo não Codificante/metabolismo , Animais , Linhagem Celular Tumoral , AMP Cíclico/biossíntese , Feminino , Humanos , Camundongos Endogâmicos C57BL , RNA Longo não Codificante/genética
16.
BMC Med Genet ; 20(1): 26, 2019 01 31.
Artigo em Inglês | MEDLINE | ID: mdl-30704411

RESUMO

BACKGROUND: Schizophrenia is a severe neurodevelopmental disorder with a complex genetic and environmental etiology. Abnormal glutamate ionotropic N-methyl-D-aspartate receptor (NMDA) type subunit 1 (NR1) may be a potential cause of schizophrenia. METHODS: We conducted a case-control study to investigate the association between the GRIN1 gene, which encodes the NR1 subunit, and the risk of schizophrenia in a northern Chinese Han population using Sanger DNA sequencing. The dual luciferase reporter assay was used to detect the influence of two different haplotypes on GRIN1 gene expression. RESULTS: Seven SNPs (single nucleotide polymorphisms), including rs112421622 (- 2019 T/C), rs138961287 (- 1962--1961insT), rs117783907 (-1945G/T), rs181682830 (-1934G/A), rs7032504 (-1742C/T), rs144123109 (-1140G/A), and rs11146020 (-855G/C) were detected in the study population. Rs117783907 (-1945G/T) was associated with the occurrence of schizophrenia as a protective factor. The genotype frequencies of rs138961287 (- 1962--1961insT) and rs11146020 (-855G/C) were statistically different between cases and controls (p < 0.0083). The other four variations were not shown to be associated with the disease. Two haplotypes were composed of the seven SNPs, and distribution of T-del-G-G-C-G-G was significantly different between the case and control groups. However, the dual luciferase reporter assay showed that neither of the haplotypes affected luciferase expression in HEK-293 and SK-N-SH cell lines. CONCLUSIONS: The GRIN1 gene may be related to the occurrence of schizophrenia. Additional research will be needed to fully ascertain the role of GRIN1 in the etiology of schizophrenia.


Assuntos
Grupo com Ancestrais do Continente Asiático/etnologia , Proteínas do Tecido Nervoso/genética , Polimorfismo de Nucleotídeo Único , Receptores de N-Metil-D-Aspartato/genética , Esquizofrenia/genética , Análise de Sequência de DNA/métodos , Adulto , Grupo com Ancestrais do Continente Asiático/genética , Estudos de Casos e Controles , Feminino , Frequência do Gene , Estudos de Associação Genética , Predisposição Genética para Doença , Células HEK293 , Haplótipos , Humanos , Masculino , Pessoa de Meia-Idade , Elementos Reguladores de Transcrição , Esquizofrenia/etnologia
17.
Electrophoresis ; 40(11): 1591-1599, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30740746

RESUMO

Semi-nested PCR with allele-specific (AS) primers and sequencing of mitochondrial DNA (mtDNA) were performed to analyze and interpret DNA mixtures, especially when biological materials were degraded or contained a limited amount of DNA. SNP-STR markers were available to identify the minor DNA component using AS-PCR; moreover, SNPs in mtDNA could be used when the degraded or limited amounts of DNA mixtures were not successful with SNP-STR markers. Five pairs of allele-specific primers were designed based on three SNPs (G15043A, T16362C, and T16519C). The sequence of mtDNA control region of minor components was obtained using AS-PCR and sequencing. Sequences of the amplification fragments were aligned and compared with the sequences of known suspects or databases. When this assay was used with the T16362C and T16519C SNPs, we found it to be highly sensitive for detecting small amounts of DNA (∼30 pg) and analyzing DNA mixtures of two contributors, even at an approximately 1‰ ratio of minor and major components. An exception was tests based on the SNP G15043A, which required approximately 300 pg of a 1% DNA mixture. In simulated three contributor DNA mixtures (at rate of 1:1:1), control region fragments from each contributor were detected and interpreted. AS-PCR combined with semi-nested PCR was successfully used to identify the mtDNA control region of each contributor, providing biological evidence for excluding suspects in forensic cases, especially when biological materials were degraded or had a limited amount of DNA.

18.
Front Mol Neurosci ; 12: 13, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30766477

RESUMO

Abnormal expression of the 5-HT1A receptor, which is encoded by the HTR1A gene, leads to susceptibilities to neuropsychiatric disorders such as depression, anxiety, and schizophrenia. miRNAs regulate gene expression by recognizing the 3'-UTR region of mRNA. This study evaluated the miRNAs that might identify and subsequently determine the regulatory mechanism of HTR1A gene. Using the HEK-293, U87, SK-N-SH and SH-SY5Y cell lines, we determined the functional sequence of the 3'-UTR region of the HTR1A gene and predicted miRNA binding. Dual luciferase reporter assay and Western Blot were used to confirm the effect of miRNA mimics and inhibitors on endogenous 5-HT1A receptors. In all cell lines, gene expression of the -17 bp to +443 bp fragment containing the complete sequence of the 3'-UTR region was significantly decreased, although mRNA quantification was not different. The +375 bp to +443 bp sequence, which exhibited the most significant change in relative chemiluminescence intensity, was recognized by hsa-miR-3177-5p and hsa-miR-3178. In HEK-293 and U87 cells, hsa-miR-3177-5p significantly inhibited the 5-HT1A receptor expression, while a hsa-miR-3178 inhibitor up-regulated HTR1A gene expression in SK-N-SH and SH-SY5Y cells. By constructing the pmirGLO-vector with the mutated HTR1A gene, we further confirmed that hsa-miR-3177-5p recognized the HTR1A gene tgtacaca at +377 bp to +384 bp, and the +392 bp to +399 bp fragment cgcgccca was identified by hsa-miR-3178. hsa-miR-3177-5p and hsa-miR-3178 had significant inhibitory effects on expression of the HTR1A gene and 5-HT1A receptor and may directly participate in the development of neuropsychiatric diseases.

19.
Mikrochim Acta ; 186(3): 193, 2019 02 18.
Artigo em Inglês | MEDLINE | ID: mdl-30778686

RESUMO

Pyrenylbutyric acid and streptavidin were coupled to films of reduced graphene oxide (rGO) and then conjugated to a biotinylated broad-spectrum monoclonal antibody against aflatoxins (AFs). It is shown that such films can efficiently and selectively capture AFs inculding AFB1, AFB2, AFG1, AFG2, AFM1 and AFM2. The rGO films were characterized by using scanning electron microscopy, energy-dispersive spectroscopy, and raman spectroscopy. The selectivity and purification performance of the antibody-loaded rGO films were investigated. They were applied to the purification of extremely small samples (100 µL) of AFs-spiked rabbit serum after enzymatic hydrolysis. The AFs were analyzed by ultra-performance liquid chromatography coupled to tandem mass spectrometry. The limits of detection for the six AFs investigated ranged from 50 to 170 pg·mL-1. The average recoveries of AFs in spiked rabbit serum samples ranged from 55% to 75%, with relative standard deviations of less than 9.4%. Graphical abstract Design of a multifunctional sandwich film that consists of a reduced graphene oxide film base, a pyrenylbutyric acid middle layer and a broad-specificity anti-AF monoclonal antibody surface layer. It was successfully applied to the determination of aflatoxins in only 100 µL of rabbit serum samples with satisfactory selectivity and acceptable accuracy.


Assuntos
Aflatoxinas/sangue , Anticorpos Monoclonais/química , Grafite/química , Animais , Técnicas Biossensoriais , Ácido Butírico/química , Cromatografia de Afinidade , Cromatografia Líquida de Alta Pressão , Oxirredução , Pirenos/química , Coelhos , Sensibilidade e Especificidade , Estreptavidina/química , Propriedades de Superfície , Espectrometria de Massas em Tandem
20.
Mol Oncol ; 13(4): 959-977, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30714292

RESUMO

Breast cancer susceptibility gene 1 (BRCA1) has been implicated in modulating metabolism via transcriptional regulation. However, direct metabolic targets of BRCA1 and the underlying regulatory mechanisms are still unknown. Here, we identified several metabolic genes, including the gene which encodes glutamate-oxaloacetate transaminase 2 (GOT2), a key enzyme for aspartate biosynthesis, which are repressed by BRCA1. We report that BRCA1 forms a co-repressor complex with ZBRK1 that coordinately represses GOT2 expression via a ZBRK1 recognition element in the promoter of GOT2. Impairment of this complex results in upregulation of GOT2, which in turn increases aspartate and alpha ketoglutarate production, leading to rapid cell proliferation of breast cancer cells. Importantly, we found that GOT2 can serve as an independent prognostic factor for overall survival and disease-free survival of patients with breast cancer, especially triple-negative breast cancer. Interestingly, we also demonstrated that GOT2 overexpression sensitized breast cancer cells to methotrexate, suggesting a promising precision therapeutic strategy for breast cancer treatment. In summary, our findings reveal that BRCA1 modulates aspartate biosynthesis through transcriptional repression of GOT2, and provides a biological basis for treatment choices in breast cancer.


Assuntos
Aspartato Aminotransferase Mitocondrial/genética , Ácido Aspártico/biossíntese , Proteína BRCA1/metabolismo , Regiões Promotoras Genéticas , Proteínas Repressoras/metabolismo , Animais , Aspartato Aminotransferase Mitocondrial/metabolismo , Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Células HEK293 , Humanos , Ácidos Cetoglutáricos/metabolismo , Metotrexato/farmacologia , Camundongos , Pessoa de Meia-Idade , Modelos Biológicos , Fenótipo , Ligação Proteica/efeitos dos fármacos , Transcrição Genética/efeitos dos fármacos
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