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1.
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.

2.
Genes (Basel) ; 10(10)2019 10 12.
Artigo em Inglês | MEDLINE | ID: mdl-31614865

RESUMO

This study identified a transcription factor that might bind to the 5' regulatory region of the HTR1A and explored the potential effect on 5-HT1A receptor expression. Based on JASPAR predictions, the binding of the transcription factor was demonstrated using the electrophoretic mobility shift assay (EMSA). Vectors over-expressing the transcription factor were co-transfected into HEK-293 and SK-N-SH cells with the recombinant pGL3 vector, and relative fluorescence intensity was measured to determine regulatory activity. Additionally, the qRT-PCR and Western blot were also used to identify whether the transcription factor modulated the endogenous expression of 5-HT1A receptor. The results suggest that the transcription factor CCAA/T enhancer binding protein beta (CEBPB) likely binds to the -1219 to -1209 bp (ATG+1) region of the HTR1A. Two sequences located in the -722 to -372 bp and -119 to +99 bp were also identified. Although the effect of CEBPB on endogenous 5-HT1A receptor expression was not significant, it exhibited the strong inhibition on the relative fluorescence intensity and the mRNA level of HTR1A. CEBPB inhibited the human HTR1A expression by binding to the sequence -1219 - -1209 bp. This is useful and informative for ascertaining the regulation of 5-HT1A receptor and mental diseases.

3.
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.

4.
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.

6.
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.

7.
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.

8.
Neuropsychiatr Dis Treat ; 15: 143-155, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30643413

RESUMO

Background: In order to explore the association between the SLC6A4 gene and the risk of schizophrenia, an updated meta-analysis was conducted using a total of 46 scientific articles. Methods: Through a literature search, papers studied included 35 articles on serotonin-transporter-linked polymorphic region (5-HTTLPR) with 8,752 cases and 10,610 controls, 17 articles on second intron variable number of tandem repeats with 7,284 cases and 8,544 controls, four studies on rs1042173 with 1,351 cases and 2,101 controls, and four studies on rs140700 with 1,770 cases and 2,386 controls. Pooled, subgroup, and sensitivity analyses were performed, and the results were visualized by forest and funnel plots. Results: An association between 5-HTTLPR and the risk of schizophrenia was not found, except for an Indian subgroup analysis (Pz =0.014, OR =1.749, 95% CI =1.120-2.731). A 10 repeats/12 repeats (10R/12R) genotype was a protective factor against schizophrenia (Pz =0.020, OR =0.789, 95% CI =0.646-0.963), but a 12R/12R genotype was a risk factor for schizophrenia (Pz =0.004, OR =1.936, 95% CI =1.238-3.029) in the pooled analyses. In Caucasians, a GG genotype of rs1042173 may be a risk factor for schizophrenia (Pz =0.006, OR =1.299, 95% CI =1.079-1.565). No association was found between rs140700 and the risk for schizophrenia. Conclusion: Through meta-analysis, we were able to gain insight into previously reported associations between SLC6A4 polymorphism and schizophrenia.

9.
BMC Genet ; 19(1): 115, 2018 12 29.
Artigo em Inglês | MEDLINE | ID: mdl-30594152

RESUMO

BACKGROUND: The serotonin neurotransmitter (5-HT) and its receptors have important roles in neuropsychiatric disorders such as schizophrenia. The aim of this study was to investigate the functional sequences of the 5' regulation region of the human HTR1A gene to explore the effects on the expression of the 5-HT1A receptor. METHODS: Fourteen recombinant pGL3-basic vectors containing deletion fragments of the HTR1A gene regulatory region were transfected with HEK-293 and SK-N-SH cells. The relative chemiluminescence intensities of different length fragments were analyzed. The JASPAR software was used for the prediction of transcription factors. RESULTS: In the HEK-293 cells, the relative chemiluminescence intensity of the - 1649 bp to - 1550 bp (ATG + 1) fragment was significantly different. Two inhibitory activity regions were found in the - 1409 bp to - 1381 bp and - 1196 bp to - 1124 bp fragments, which might be bound to the GATA or SOX10 transcription factors as predicted by the JASPAR software. In addition, the fragments located from - 1124 bp to - 1064 bp and from - 908 bp to - 722 bp up-regulated protein expression. Only the sequence from - 1550 bp to - 1409 bp demonstrated a difference in luciferase expression in the both cell lines. According to the results of the 5'-UTR truncated vectors, there was a repression region at the distal end of the 5'-UTR, an enhancer region might be present at the proximal end of the transcription start site. CONCLUSIONS: Although the functional sequences of the HTR1A gene regulatory region were confirmed, the regulatory factors and functional components require further investigation.


Assuntos
Receptor 5-HT1A de Serotonina/genética , Regiões 5' não Traduzidas , Linhagem Celular Tumoral , Regulação da Expressão Gênica , Genes Reporter , Células HEK293 , Humanos , Receptor 5-HT1A de Serotonina/metabolismo , Elementos Reguladores de Transcrição , Esquizofrenia/genética , Esquizofrenia/patologia , Deleção de Sequência , Fatores de Transcrição/química , Fatores de Transcrição/metabolismo , Transfecção
10.
J Mol Neurosci ; 65(4): 438-443, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30022436

RESUMO

Dysregulation of dopamine receptor D1 (DRD1) is involved in multiple neuropsychiatric disorders. The 5' regulatory region of DRD1 has not been characterized fully. We applied the luciferase assay and the electrophoretic mobility shift assay to explore the activity of the 5' regulatory region of DRD1 in SH-SY5Y and 293T cells. We found that the promoter region of DRD1 corresponded to positions - 1250 to + 250 in the DNA sequence, and the putative core promoter region was from - 113 to + 250 (transcriptional start site of exon, +1). The sequence 5'-gggacgcgcgggcggggtgggctgtgccccgcgggaaccccgccggcctgtgcgcttgctg-3' was identified as a possible transcription factor-binding domain. Further research is warranted to explore the function of the 5' regulatory region of DRD1.


Assuntos
Regiões Promotoras Genéticas , Receptores de Dopamina D1/genética , Linhagem Celular Tumoral , Células HEK293 , Humanos , Motivos de Nucleotídeos , Ligação Proteica , Receptores de Dopamina D1/metabolismo , Fatores de Transcrição
11.
Neuropsychiatr Dis Treat ; 14: 153-164, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29379288

RESUMO

To explore the association between DRD4 polymorphisms and schizophrenia risk, a meta-analysis was carried out with 41 case-control articles. Specifically, we included 28 articles (5,735 cases and 5,278 controls) that pertained to the 48 bp variable number tandem repeat (VNTR) polymorphism, nine articles (1,517 cases and 1,746 controls) that corresponded to the 12 bp tandem repeat (TR), six articles (1,912 cases and 1,836 controls) that addressed the 120 bp TR, 10 articles (2,927 cases and 2,938 controls) that entailed the -521 C>T polymorphism, six articles (1,735 cases and 1,724 controls) that pertained to the -616 C>G polymorphism, and four articles (1,191 cases and 1,215 controls) that involved the -376 C>T polymorphism. Pooled analysis, subgroup analysis, and sensitivity analysis were performed, and the data were visualized by means of forest and funnel plots. Results of pooled analysis indicated that the -521 CC variant (Pz=0.009, odds ratio [OR] =1.218, 95% confidence interval [CI] =1.050-1.413) and genotype L/L (ie, long allele) of the 120 bp TR were risk factors of schizophrenia (Pz=0.004, OR =1.275, 95% CI =1.081-1.504). The 48 bp VNTR, the 12 bp TR, the -616 C>G polymorphism, and the -376 C>T polymorphism were not associated with schizophrenia. Additional research is warranted to explore the association between polymorphisms of DRD4 and schizophrenia risk.

12.
Mitochondrial DNA A DNA Mapp Seq Anal ; 29(2): 250-255, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-28093929

RESUMO

This study characterized the genetic variations of mitochondrial DNA (mtDNA) to elucidate the maternal genetic structure of Liaoning Han Chinese. A total of 317 blood samples of unrelated individuals were collected for analysis in Liaoning Province. The mtDNA samples were analyzed using two distinct methods: sequencing of the hypervariable sequences I and II (HVSI and HVSII), and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the coding region. The results indicated a high gene diversity value (0.9997 ± 0.0003), a high polymorphism information content (0.99668) and a random match probability (0.00332). These samples were classified into 305 haplotypes, with 9 shared haplotypes. The most common haplogroup was D4 (12.93%). The principal component analysis map, the phylogenetic tree map, and the genetic distance matrix all indicated that the genetic distance of the Liaoning Han population from the Tibetan group was distant, whereas that from the Miao group was relatively close.


Assuntos
Grupo com Ancestrais do Continente Asiático/etnologia , Grupo com Ancestrais do Continente Asiático/genética , DNA Mitocondrial/genética , Polimorfismo de Nucleotídeo Único , China/etnologia , Feminino , Genética Populacional , Haplótipos , Humanos , Masculino , Herança Materna , Filogenia , Polimorfismo de Fragmento de Restrição , Análise de Componente Principal
13.
PLoS One ; 12(8): e0182769, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28846698

RESUMO

To determine whether mitochondrial DNA (mtDNA) variations are associated with schizophrenia, 313 patients with schizophrenia and 326 unaffected participants of the northern Chinese Han population were included in a prospective study. Single-nucleotide polymorphisms (SNPs) including C5178A, A10398G, G13708A, and C13928G were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Hypervariable regions I and II (HVSI and HVSII) were analyzed by sequencing. The results showed that the 4 SNPs and 11 haplotypes, composed of the 4 SNPs, did not differ significantly between patient and control groups. No significant association between haplogroups and the risk of schizophrenia was ascertained after Bonferroni correction. Drawing a conclusion, there was no evidence of an association between mtDNA (the 4 SNPs and the control region) and schizophrenia in the northern Chinese Han population.


Assuntos
DNA Mitocondrial/genética , Predisposição Genética para Doença , Polimorfismo de Nucleotídeo Único , Esquizofrenia/genética , Adulto , Grupo com Ancestrais do Continente Asiático/genética , China , Feminino , Estudos de Associação Genética , Haplótipos , Humanos , Masculino , Pessoa de Meia-Idade
14.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 25(2): 393-397, 2017 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-28446281

RESUMO

OBJECTIVE: To study the effect of cyclin dependent kinase(CDK) inhibitor LS-007 on acute lymphoblastic leukemia and its mechanism. METHODS: The acute lymphocytic leukemia cell line was cultured and treated by LS-007, flavopiridol and ABT-199, then the changes of apoptosis-related factor mRNA and protein levels were detected by using mRNA quantitative PCR and Werstern blot. RESULTS: quantitative PCR and Western blot detection showed that the levels of antiapoptotic protein decreased significantly in acute lymphoblastic leukemia cells after LS-007 treatment, and the pro-apoptotic effect of LS-007 combined with ABT-199 was much better. CONCLUSION: LS-007 can affect the phosphorylation of RNA polymerase sites and promote cell apoptosis through changing the activities of CDK, thus having some positive significance for relieving acute lymphoblastic leukemia.


Assuntos
Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamento farmacológico , Pirimidinas/farmacologia , Sulfonamidas/farmacologia , Apoptose , Linhagem Celular Tumoral , Quinases Ciclina-Dependentes , Humanos , Células Tumorais Cultivadas
15.
Neurosci Lett ; 638: 51-54, 2017 01 18.
Artigo em Inglês | MEDLINE | ID: mdl-27939355

RESUMO

The aim of this study was to explore the role of two SNPs (rs6295 & rs113195492) in the HTR1A gene promoter region that regulates the expression of the 5-HT1A receptor. A fragment spanning from -551 to +672 of HTR1A (Transcription start site +1) was cloned into the pGL-3 Basic Vector and three haplotype plasmids composed of two SNPs were constructed. HEK-293 cells and the SK-N-SH cells were transfected with the three plasmids, and the relative fluorescence intensity was measured. In HEK-293 cells, there was a significant difference when the relative fluorescence intensity of plasmid 1-1 was compared to that of plasmid 2-1. However, no significant difference was observed when the luciferase expression of plasmid 2-1 and plasmid 3-1 was analyzed. We also found that the expression trend of the SK-N-SH cells was similar to the HEK-293 cells, but the overall relative fluorescence intensity of the SK-N-SH cells was lower than that of the HEK-293 cells. Our finding showed that the rs6295 SNP, as a suspected variant that indicates susceptibility to schizophrenia, exhibited a higher transcriptional activity. The influence of the rs113195492 locus on schizophrenia needs to be explored further.


Assuntos
Receptor 5-HT1A de Serotonina/genética , Linhagem Celular , Haplótipos , Humanos , Polimorfismo de Nucleotídeo Único , Regiões Promotoras Genéticas , Receptor 5-HT1A de Serotonina/metabolismo , Transcrição Genética
17.
Zhongguo Yi Liao Qi Xie Za Zhi ; 30(3): 199-205, 2006 May.
Artigo em Chinês | MEDLINE | ID: mdl-16929780

RESUMO

This article makes a pilot study on the key points of the quality management system of in-vitro diagnostic reagents by analyzing the technical characteristics and production methods of these products as well as the status in quo, and problems the in-vitro diagnostic reagent industry in China is facing nowadays. It can serve as a reference to the supervision departments and the manufacturers in this field which are establishing and running the quality management system.


Assuntos
Indicadores e Reagentes , Garantia da Qualidade dos Cuidados de Saúde/organização & administração , Tecnologia Farmacêutica/organização & administração , China , Equipamentos e Provisões/normas , Humanos , Indicadores e Reagentes/química , Indicadores e Reagentes/normas , Projetos Piloto , Controle de Qualidade , Kit de Reagentes para Diagnóstico/normas , Gestão da Segurança , Tecnologia Farmacêutica/normas , Gestão da Qualidade Total
18.
Zhongguo Yi Liao Qi Xie Za Zhi ; 30(2): 123-8, 2006 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-16830807

RESUMO

Safety and effectiveness of medical devices is a focus of people's attention, which is also a central work for the government's administration. Carrying out safety standards of medical devices, implementing the quality system standards and risk management standards are supplemented with each other. The risk management is an indispensable part to the quality management system of medical devices.


Assuntos
Segurança de Equipamentos/normas , Equipamentos e Provisões/normas , Gestão de Riscos , Análise de Falha de Equipamento , Segurança de Equipamentos/métodos , Humanos , Vigilância de Produtos Comercializados/métodos , Medição de Risco , Fatores de Risco , Gestão da Segurança
19.
Zhongguo Yi Liao Qi Xie Za Zhi ; 29(2): 124-30, 2005 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-16011119

RESUMO

This article introduces the definition, classification, premarket admission and other administering specialities about In-Vitro Diagnostic Reagents in the U.S.A. and China. And by analyzing manufacture and administration of In-Vitro Diagnostic Reagents in our country, It is pointed out that a suitable administering model in accordance with the characteristics of In-Vitro Diagnostic Reagents should be adopted to perfect the administration.


Assuntos
Kit de Reagentes para Diagnóstico , China , Aprovação de Equipamentos , Indicadores e Reagentes/classificação , Indicadores e Reagentes/normas , Controle de Qualidade , Kit de Reagentes para Diagnóstico/classificação , Kit de Reagentes para Diagnóstico/normas , Estados Unidos , United States Food and Drug Administration
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