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1.
Lab Invest ; 104(4): 100324, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38220044

RESUMO

Meningiomas rank among the most common intracranial tumors, and surgery stands as the primary treatment modality for meningiomas. The precise subtyping and diagnosis of meningiomas, both before and during surgery, play a pivotal role in enabling neurosurgeons choose the optimal surgical program. In this study, we utilized multiphoton microscopy (MPM) based on 2-photon excited fluorescence and second-harmonic generation to identify 5 common meningioma subtypes. The morphological features of these subtypes were depicted using the MPM multichannel mode. Additionally, we developed 2 distinct programs to quantify collagen content and blood vessel density. Furthermore, the lambda mode of the MPM characterized architectural and spectral features, from which 3 quantitative indicators were extracted. Moreover, we employed machine learning to differentiate meningioma subtypes automatically, achieving high classification accuracy. These findings demonstrate the potential of MPM as a noninvasive diagnostic tool for meningioma subtyping and diagnosis, offering improved accuracy and resolution compared with traditional methods.


Assuntos
Neoplasias Meníngeas , Meningioma , Humanos , Meningioma/diagnóstico por imagem , Colágeno , Microscopia de Fluorescência por Excitação Multifotônica/métodos , Neoplasias Meníngeas/diagnóstico por imagem , Computadores
2.
Plant Cell ; 33(4): 1212-1228, 2021 05 31.
Artigo em Inglês | MEDLINE | ID: mdl-33693937

RESUMO

Panicle size and grain number are important agronomic traits and influence grain yield in rice (Oryza sativa), but the molecular and genetic mechanisms underlying panicle size and grain number control remain largely unknown in crops. Here we report that LARGE2 encodes a HECT-domain E3 ubiquitin ligase OsUPL2 and regulates panicle size and grain number in rice. The loss of function large2 mutants produce large panicles with increased grain number, wide grains and leaves, and thick culms. LARGE2 regulates panicle size and grain number by repressing meristematic activity. LARGE2 is highly expressed in young panicles and grains. Biochemical analyses show that LARGE2 physically associates with ABERRANT PANICLE ORGANIZATION1 (APO1) and APO2, two positive regulators of panicle size and grain number, and modulates their stabilities. Genetic analyses support that LARGE2 functions with APO1 and APO2 in a common pathway to regulate panicle size and grain number. These findings reveal a novel genetic and molecular mechanism of the LARGE2-APO1/APO2 module-mediated control of panicle size and grain number in rice, suggesting that this module is a promising target for improving panicle size and grain number in crops.


Assuntos
Oryza/fisiologia , Proteínas de Plantas/genética , Sementes/genética , Ubiquitina-Proteína Ligases/genética , Clonagem Molecular , Produtos Agrícolas/genética , Regulação da Expressão Gênica de Plantas , Redes Reguladoras de Genes , Mutação , Oryza/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Estabilidade Proteica , Ubiquitina-Proteína Ligases/metabolismo
3.
Cell Commun Signal ; 22(1): 302, 2024 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-38831335

RESUMO

The ubiquitination-mediated protein degradation exerts a vital role in the progression of multiple tumors. NEDD4L, which belongs to the E3 ubiquitin ligase NEDD4 family, is related to tumor genesis, metastasis and drug resistance. However, the anti-tumor role of NEDD4L in esophageal carcinoma, and the potential specific recognition substrate remain unclear. Based on public esophageal carcinoma database and clinical sample data, it was discovered in this study that the expression of NEDD4L in esophageal carcinoma was apparently lower than that in atypical hyperplastic esophageal tissue and esophageal squamous epithelium. Besides, patients with high expression of NEDD4L in esophageal carcinoma tissue had longer progression-free survival than those with low expression. Experiments in vivo and in vitro also verified that NEDD4L suppressed the growth and metastasis of esophageal carcinoma. Based on co-immunoprecipitation and proteome analysis, the NEDD4L ubiquitination-degraded protein ITGB4 was obtained. In terms of the mechanism, the HECT domain of NEDD4L specifically bound to the Galx-ß domain of ITGB4, which modified the K915 site of ITGB4 in an ubiquitination manner, and promoted the ubiquitination degradation of ITGB4, thus suppressing the malignant phenotype of esophageal carcinoma.


Assuntos
Progressão da Doença , Neoplasias Esofágicas , Integrina beta4 , Ubiquitina-Proteína Ligases Nedd4 , Proteólise , Ubiquitinação , Neoplasias Esofágicas/patologia , Neoplasias Esofágicas/metabolismo , Neoplasias Esofágicas/genética , Humanos , Ubiquitina-Proteína Ligases Nedd4/metabolismo , Ubiquitina-Proteína Ligases Nedd4/genética , Animais , Linhagem Celular Tumoral , Integrina beta4/metabolismo , Integrina beta4/genética , Camundongos Nus , Camundongos , Proliferação de Células , Masculino , Regulação Neoplásica da Expressão Gênica , Feminino
4.
Carcinogenesis ; 43(7): 624-634, 2022 08 30.
Artigo em Inglês | MEDLINE | ID: mdl-35605971

RESUMO

Proteasome dysregulation is a common feature of cancer and a critical risk for tumorigenesis. However, the characteristics of proteasome components in tumor development and metastasis are poorly understood. PSMA5, an α5 subunit of the 20S core proteasome, is associated with the degradation of intracellular proteins. Increasing evidence indicated that it is involved in tumor development, but the underlying mechanism has remained unknown. Here, we show that PSMA5 is upregulated in lung adenocarcinoma (LUAD) cells and clinical LUAD tissues. Moreover, its upregulation is positively associated with lymph node metastasis and the poor prognosis of LUAD patients. PSMA5 knockdown inhibited the proliferation, invasion and metastasis of LUAD cells in vitro and in vivo, induced apoptosis of LUAD cells and sensitized LUAD cells to cisplatin. Furthermore, investigations revealed that PSMA5 overexpression inhibited cell apoptosis by activating the janus kinase (JAK)/signal transducer and activator of transcription (STAT) signaling pathway in LUAD cells. In total, our results demonstrate that PSMA5 may function as a prognostic factor in LUAD. In addition, PSMA5 is a promising therapeutic target for LUAD, as its depletion induces cell apoptosis by inhibiting the JAK/STAT pathway.


Assuntos
Adenocarcinoma de Pulmão , Neoplasias Pulmonares , Complexo de Endopeptidases do Proteassoma , Adenocarcinoma de Pulmão/genética , Adenocarcinoma de Pulmão/patologia , Linhagem Celular Tumoral , Proliferação de Células/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Janus Quinases/genética , Janus Quinases/metabolismo , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Complexo de Endopeptidases do Proteassoma/genética , Complexo de Endopeptidases do Proteassoma/metabolismo , Fatores de Transcrição STAT/metabolismo , Transdução de Sinais
5.
Gastrointest Endosc ; 95(2): 249-257, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-34371004

RESUMO

BACKGROUND AND AIMS: Lugol's iodine chromoendoscopy is an important method to detect esophageal squamous cell carcinoma. Sodium thiosulfate solution (STS) has been used to neutralize iodine after Lugol's chromoendoscopy; however, it is not available in many medical centers. The aim of the current study was to assess the efficacy of N-acetylcysteine solution (NAC) for relieving symptoms caused by Lugol's iodine chromoendoscopy. METHODS: Patients were randomized to receive either STS or NAC after spraying Lugol's iodine solution on the esophagus. The neutralizing effects for residual iodine in the esophagus and gastric mucous pool were observed. The primary endpoint was the intensity of retrosternal pain and/or heartburn measured by a visual analog scale (VAS) score 30 minutes after chromoendoscopy. Secondary endpoints were the rate of patients with any adverse symptom, rate of moderate to severe retrosternal discomfort occurring, and heart rate variability between time points before and after chromoendoscopy. RESULTS: The neutralization rates for residual iodine between the NAC and STS groups were not significantly different (P > .999). The difference of median VAS scores between the NAC and STS groups 30 minutes after chromoendoscopy was .0 (P = .719; 95% confidence interval, .0-.0), and the 95% confidence interval higher limit was .0, which was less than our prespecified margin of .5, concluding an noninferiority of NAC with regard to STS. There was no significant difference between the 2 groups regarding the rate of patients with any adverse symptom, rate of moderate to severe retrosternal discomfort, or heart rate variability at 5 minutes or 30 minutes after chromoendoscopy. CONCLUSION: As a very easily accessible reagent in clinical circumstances, NAC can also alleviate mucosal irritation symptoms induced by Lugol's chromoendoscopy at similar efficacy as STS and can be routinely recommended. (Clinical trial registration number: NCT04764643.).


Assuntos
Neoplasias Esofágicas , Carcinoma de Células Escamosas do Esôfago , Acetilcisteína/uso terapêutico , Corantes , Esofagoscopia/métodos , Humanos , Iodetos/uso terapêutico , Tiossulfatos
6.
J Cell Mol Med ; 24(13): 7439-7450, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32515550

RESUMO

SIRT2 is a NAD+ -dependent deacetylase that deacetylates a diverse array of protein substrates and is involved in many cellular processes, including regulation of inflammation. However, its precise role in the inflammatory process has not completely been elucidated. Here, we identify heat-shock protein 90α (Hsp90α) as novel substrate of SIRT2. Functional investigation suggests that Hsp90 is deacetylated by SIRT2, such that overexpression and knock-down of SIRT2 altered the acetylation level of Hsp90. This subsequently resulted in disassociation of Hsp90 with glucocorticoid receptor (GR), and translocation of GR to the nucleus. This observation was further confirmed by glucocorticoid response element (GRE)-driven reporter assay. Nuclear translocation of GR induced by SIRT2 overexpression repressed the expression of inflammatory cytokines, which were even more prominent under lipopolysaccharide (LPS) stimulation. Conversely, SIRT2 knock-down resulted in the up-regulation of cytokine expression. Mutation analysis indicated that deacetylation of Hsp90 at K294 is critical for SIRT2-mediated regulation of cytokine expression. These data suggest that SIRT2 reduces the extent of LPS-induced inflammation by suppressing the expression of inflammatory factors via SIRT2-Hsp90-GR axis.


Assuntos
Proteínas de Choque Térmico HSP90/metabolismo , Inflamação/metabolismo , Receptores de Glucocorticoides/metabolismo , Transdução de Sinais , Sirtuína 2/metabolismo , Acetilação/efeitos dos fármacos , Sequência de Aminoácidos , Animais , Linhagem Celular , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Escherichia coli/metabolismo , Humanos , Lipopolissacarídeos/farmacologia , Modelos Biológicos , Ligação Proteica/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Interferente Pequeno/metabolismo , Ratos , Transdução de Sinais/efeitos dos fármacos , Sirtuína 2/química , Solubilidade
7.
J Cell Biochem ; 121(2): 2019-2026, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31691337

RESUMO

Acute lymphoblastic leukemia (ALL) is the most frequently diagnosed cancer in children and single-nucleotide polymorphisms (SNPs) in certain genes influence risk of ALL. Although FOXO3 had been demonstrated to be involved leukemia, the role of FOXO3 polymorphisms was still not clear. In the present study, we explored the association of FOXO3 SNPs with ALL risk in Chinese children. We genotyped four polymorphisms (rs17069665 A>G, rs4945816 T>C, rs4946936 C>T, and rs9400241 A>C) of FOXO3 in 425 ALL cases and 1339 health controls. The associations were estimated by odds ratios (ORs) with their 95% confidence intervals (CIs). Further analyses were performed to explore associations of rs17069665 and rs9400241 with ALL susceptibility in terms of age, gender, immunophenotype, minimal residual disease (MRD), and other clinical characteristics. We found rs17069665 related to the increased ALL risk (OR = 1.76; 95% CI = 1.02-3.04), rs9400241 related to decreased ALL risk (OR = 0.80; 95% CI = 0.64-0.99). The effects of rs17069665 on ALL risk were more predominant in males and children < 10 years, and patients with lower rates of platelet or neutrophil. As for rs9400241, the effects were more predominant in children < 10 years, and in patients with pre B ALL, positive MRD, anemia, or hepatomegaly. In conclusion, FOXO3 gene polymorphisms influence the risk of ALL in children and might be a potential biomarker for ALL susceptibility.


Assuntos
Povo Asiático/genética , Biomarcadores Tumorais/genética , Proteína Forkhead Box O3/genética , Predisposição Genética para Doença , Polimorfismo de Nucleotídeo Único , Leucemia-Linfoma Linfoblástico de Células Precursoras/epidemiologia , Estudos de Casos e Controles , Criança , China/epidemiologia , Feminino , Seguimentos , Genótipo , Humanos , Masculino , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/patologia , Prognóstico , Fatores de Risco
8.
Cancer Sci ; 111(8): 2824-2836, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32495982

RESUMO

In this study, we explored expression and functions of circular RNA LPAR3 (circLPAR3) in esophageal squamous cell carcinoma (ESCC). The differential expression of circular RNAs (circRNAs) in 10 ESCC and corresponding paracarcinoma tissues was analyzed through circRNA microarray, then the candidate circRNAs were detected and verified through quantitative RT-PCR, and a novel circRNA was screened, which was circLPAR3. Circular RNA LPAR3 showed apparently high expression in ESCC tissues and cells, which was closely correlated with the clinical stage and lymph node metastasis of ESCC patients. Circular RNA LPAR3 was mainly located in the cytoplasm of ESCC cells, which was more stable than the baseline gene. Circular RNA LPAR3 upregulated MET gene expression through sponge adsorption of microRNA (miR)-198, activated the RAS/MAPK and the PI3K/Akt pathways, and promoted ESCC cell migration, invasion, and metastasis in vivo and in vitro. However, it had no effect on ESCC cell proliferation. Circular RNA LPAR3 can regulate the miR-198-MET signal axis to promote the migration, invasion, and metastasis of esophageal cancer cells, which can thereby serve as a potential diagnostic and therapeutic target of esophageal cancer.


Assuntos
Neoplasias Esofágicas/genética , Carcinoma de Células Escamosas do Esôfago/genética , Neoplasias Pulmonares/genética , MicroRNAs/metabolismo , RNA Circular/metabolismo , Linhagem Celular Tumoral , Movimento Celular/genética , Neoplasias Esofágicas/patologia , Neoplasias Esofágicas/cirurgia , Carcinoma de Células Escamosas do Esôfago/patologia , Carcinoma de Células Escamosas do Esôfago/secundário , Carcinoma de Células Escamosas do Esôfago/cirurgia , Esofagectomia , Esôfago/patologia , Esôfago/cirurgia , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Humanos , Pulmão/patologia , Neoplasias Pulmonares/secundário , Metástase Linfática/genética , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica/genética , Análise de Sequência com Séries de Oligonucleotídeos , Proteínas Proto-Oncogênicas c-met/genética , RNA Circular/genética , Receptores de Ácidos Lisofosfatídicos/genética , Ensaios Antitumorais Modelo de Xenoenxerto
9.
BMC Med Genet ; 20(1): 169, 2019 11 06.
Artigo em Inglês | MEDLINE | ID: mdl-31694554

RESUMO

BACKGROUND: Proximal symphalangism is a rare disease with multiple phenotypes including reduced proximal interphalangeal joint space, symphalangism of the 4th and/or 5th finger, as well as hearing loss. At present, at least two types of proximal symphalangism have been identified in the clinic. One is proximal symphalangism-1A (SYM1A), which is caused by genetic variants in Noggin (NOG), another is proximal symphalangism-1B (SYM1B), which is resulted from Growth Differentiation Factor 5 (GDF5) mutations. CASE PRESENTATION: Here, we reported a Chinese family with symphalangism of the 4th and/or 5th finger and moderate deafness. The proband was a 13-year-old girl with normal intelligence but symphalangism of the 4th finger in the left hand and moderate deafness. Hearing testing and inner ear CT scan suggested that the proband suffered from structural deafness. Family history investigation found that her father (II-3) and grandmother (I-2) also suffered from hearing loss and symphalangism. Target sequencing identified a novel heterozygous NOG mutation, c.690C > G/p.C230W, which was the genetic lesion of the affected family. Bioinformatics analysis and public databases filtering further confirmed the pathogenicity of the novel mutation. Furthermore, we assisted the family to deliver a baby girl who did not carry the mutation by genetic counseling and prenatal diagnosis using amniotic fluid DNA sequencing. CONCLUSION: In this study, we identified a novel NOG mutation (c.690C > G/p.C230W) by target sequencing and helped the family to deliver a baby who did not carry the mutation. Our study expanded the spectrum of NOG mutations and contributed to genetic diagnosis and counseling of families with SYM1A.


Assuntos
Proteínas de Transporte/genética , Articulações dos Dedos/anormalidades , Artropatias/congênito , Mutação , Adolescente , Adulto , Surdez/genética , Feminino , Humanos , Artropatias/diagnóstico , Artropatias/genética , Masculino , Pessoa de Meia-Idade , Linhagem , Diagnóstico Pré-Natal
10.
Future Oncol ; 15(14): 1605-1615, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31038356

RESUMO

Aim: WASF3 has been shown to be required for invasion and metastasis in different cancers, this study is to explore the prognostic value of WASF3 in gastric cancer. Materials & methods: The coexpression of WASF3 and E-cadherin in gastric cancer patients and cells were evaluated. Results: WASF3 was overexpressed and the expression of E-cadherin was decreased in gastric cancer tissues compared with normal tissues (p < 0.001). WASF3 expression is associated with decreased expression of E-cadherin (p = 0.002). Patients with WASF3-positive expression had a poorer prognosis. The multivariate analysis showed that WASF3 expression is an independent prognostic factor related to overall survival (p = 0.027). Conclusion: Our analysis demonstrates that WASF3 expression correlates with poor outcomes and is a potential prognostic factor in gastric cancer patients.


Assuntos
Regulação Neoplásica da Expressão Gênica , Neoplasias Gástricas/genética , Neoplasias Gástricas/mortalidade , Família de Proteínas da Síndrome de Wiskott-Aldrich/genética , Adulto , Idoso , Biomarcadores Tumorais , Caderinas/genética , Caderinas/metabolismo , Linhagem Celular Tumoral , Feminino , Humanos , Imuno-Histoquímica , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Gradação de Tumores , Metástase Neoplásica , Estadiamento de Neoplasias , Prognóstico , Neoplasias Gástricas/diagnóstico , Família de Proteínas da Síndrome de Wiskott-Aldrich/metabolismo
11.
Exp Cell Res ; 369(2): 234-242, 2018 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-29802838

RESUMO

PTEN, a well-known tumor suppressor, dephosphorylates PIP3 and inhibits AKT activity. A translational variant of PTEN has been identified and termed PTEN-Long (PTEN-L). The additional 173 amino acids (PTEN-L leader) at the N-terminal constitute a potential signal peptide. Differing from canonical PTEN, PTEN-L is secreted into the extracellular fluid and re-enters recipient cells, playing the similar roles as PTEN in vivo and in vitro. This character confers the PTEN-L a therapeutic ability via directly protein delivering instead of traditional DNA and RNA vector options. In the present study, we employed PTEN-L leader to assemble a fusion protein, PTEN-L-p53, inosculated with the transcriptional regulator TP53, which is another powerful tumor suppressor. We overexpressed PTEN-L-p53 in HEK293T cells and detected it in both the cytoplasm and nucleus. Subsequently, we found that PTEN-L-p53 was secreted outside of the cells and detected in the culture media by immunoblotting. Furthermore, we demonstrated that PTEN-L-p53 freely entered the cells and suppressed the viability of U251cells (p53R273H, a cell line with p53 R273H-mutation). PTEN-L-p53 is composed of endogenous protein/peptide bearing low immunogenicity, and only the junction region between PTEN-L leader and p53 can act as a new immune epitope. Accordingly, this fusion protein can potentially be used as a therapeutic option for TP53-abnormality cancers.


Assuntos
PTEN Fosfo-Hidrolase/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Substituição de Aminoácidos , Linhagem Celular Tumoral , Movimento Celular/genética , Movimento Celular/imunologia , Movimento Celular/fisiologia , Proliferação de Células/genética , Proliferação de Células/fisiologia , Sistemas de Liberação de Medicamentos , Células HEK293 , Humanos , Mutação de Sentido Incorreto , Invasividade Neoplásica/genética , Invasividade Neoplásica/imunologia , Invasividade Neoplásica/fisiopatologia , PTEN Fosfo-Hidrolase/genética , PTEN Fosfo-Hidrolase/imunologia , Sinais Direcionadores de Proteínas/genética , Sinais Direcionadores de Proteínas/fisiologia , Transporte Proteico , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/metabolismo , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/imunologia
12.
Reprod Fertil Dev ; 31(4): 671-682, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30423284

RESUMO

Non-obstructive azoospermia (NOA) is the most severe clinical diagnosis in cases of male infertility. Although in some cases of NOA spermatozoa can be retrieved by microdissection testicular sperm extraction (micro-TESE) to fertilise eggs through intracytoplasmic sperm injection (ICSI), there remains a lack of potential biomarkers for non-invasive diagnosis before micro-TESE surgery. To determine predictive biomarkers for successful sperm retrieval before micro-TESE, the aim of this study was to explore whether microRNAs (miRNAs) were differentially expressed in testicular tissues in NOA patients in whom sperm retrieval had been successful (SSR) versus those in whom it had been unsuccessful (USR) using next-generation small RNA sequencing (RNA-Seq). In all, 180 miRNAs were identified with significantly altered expression levels between SSR and USR testicular tissues. Of these, the expression of 13 miRNAs was upregulated and that of 167 miRNAs was downregulated in the USR compared with SSR group. Unexpectedly, 86 testicular miRNAs were found to be completely absent in the USR group, but showed high expression in the SSR group, suggesting that these miRNAs may serve as biomarkers for micro-TESE and may also play an essential role in spermatogenesis. Furthermore, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses indicated that the miRNAs that differed significantly between the USR and SSR groups were involved in cell apoptosis, proliferation and differentiation, which are of considerable importance during spermatogenesis. In summary, this study identified a panel of miRNAs highly expressed in testicular tissues of SSR but not USR NOA patients, providing new insights into specific miRNAs that may play important roles in epigenetic regulation during spermatogenesis. The findings provide a basis for further elucidation of the regulatory role of miRNAs in spermatogenesis and clues to identifying useful biomarkers to predict residual spermatogenic loci in NOA patients during treatment with assisted reproductive technologies.


Assuntos
Azoospermia/metabolismo , MicroRNAs/metabolismo , Recuperação Espermática , Testículo/metabolismo , Adulto , Azoospermia/genética , Humanos , Masculino , MicroRNAs/genética , Microdissecção , Pessoa de Meia-Idade , Injeções de Esperma Intracitoplásmicas , Adulto Jovem
14.
J Assist Reprod Genet ; 35(5): 801-808, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29460056

RESUMO

PURPOSE: PIWI-interacting RNA (piRNA) is a sub-group of small RNAs about 30 nucleotides length which specifically expressed in mammalian germ cells. Although piRNAs play pivotal roles in spermatogenesis regulation, little is known in the testicular tissues of infertile men. To explore whether piRNA profile could serve as a biomarker for male infertility diagnosis in a clinic, in this study, we systematically investigated the expression profile of piRNAs in testicular tissues from the patients with non-obstructive azoospermia (NOA) between successful and unsuccessful sperm retrieval before micro-dissection testicular sperm extraction (micro-TESE). METHODS: The differential expression levels of piRNAs were evaluated using small RNA-Seq method. Ontologic analyses were performed to determine the presence of enriched biological processes. RESULTS: A total of 18,324 Homo sapiens piRNAs were identified by small RNA-Seq from NOA patient testicular tissues; among them, 959 piRNAs were significantly altered between successful and unsuccessful sperm retrieval groups, of which 951 testicular piRNAs were significantly downregulated and 8 piRNAs were upregulated in NOA patients with unsuccessful sperm retrieval (USR) groups compared to those with successful sperm retrieval (SSR) groups, respectively. Unexpectedly, 553 testicular piRNAs were found completely absent in USR but showing abundant in SSR, which suggests that those piRNAs might serve as a biomarker for micro-TESE application. A total of 20 significantly differential piRNAs (hsa-piR-20830, hsa-piR-4731, hsa-piR-6254, hsa-piR-419, hsa-piR-7152, hsa-piR-7548, hsa-piR-14195, hsa-piR-5026, hsa-piR-11482, hsa-piR-17765, hsa-piR-17102, hsa-piR-4484, hsa-piR-17260, hsa-piR-17098, hsa-piR-20511, hsa-piR-5802, hsa-piR-19121, hsa-piR-2510, hsa-piR-4745, hsa-piR-11873) were selected to further validate the RNA-Seq data by quantitative real-time polymerase chain reaction. In addition, bioinformatic analyses revealed that those altered piRNAs were involved in many important biological pathways, including apoptosis, cell proliferation, and differentiation. CONCLUSIONS: Our results demonstrate that testicular tissues from NOA patients with successful and unsuccessful spermatozoa retrieval exhibit differential piRNA profiles. This study provides a useful resource to further elucidate the regulatory role of piRNAs in spermatogenesis and provides a profound clue to identify useful biomarkers for predicting residual spermatogenic loci in NOA patients during assisted reproductive treatment.


Assuntos
Azoospermia/genética , RNA Interferente Pequeno/análise , Recuperação Espermática , Adulto , Ontologia Genética , Humanos , Masculino , Reprodutibilidade dos Testes , Análise de Sequência de RNA , Espermatogênese/genética , Transcriptoma , Resultado do Tratamento
15.
J Cell Mol Med ; 21(12): 3337-3346, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28631420

RESUMO

PTEN is a tumour suppressor that is frequently mutated in a variety of cancers. Hence, PTEN has significant potential as a therapeutic molecule. PTEN-long is an alternative translation variant, with an additional 173 amino acids added to the N-terminal of the canonical PTEN when CUG of the mRNA is utilized as the start codon. PTEN-long is secreted into serum and can re-enter cells throughout the body. One of the major barriers for gene therapy is to efficiently and specifically deliver DNA or RNA material to target cells. As an alternative approach, if a therapeutic protein can be directly delivered to target cell of interest, it should theoretically function well within the cells, particularly for genes that are deficiently expressed in vivo. Most therapeutic proteins are incapable of efficiently permeating the cell membrane. In this study, we have employed CRISPR/Cas9 gene editing tool combined with single-stranded template to edit CTG of PTEN-long to ATG in the genome. Two guide RNAs close to CTG site were found to have similar efficiency in driving PTEN-long expression. Furthermore, we detected PTEN-long expression in transfected whole-cell lysate and in concentrated culture media in Western blot. Interestingly, the culture media of PTEN-long expression can reduce Akt phosphorylation level and repress U87 cell proliferation compared to wild-type U87 or control media. Taken together, PTEN-long driven by CRISPR/Cas9 imports and exports cells and represses nearby cell proliferation, indicating the PTEN-long generated by CRISPR/Cas9 has potential to be an alternative strategy for PTEN gene therapy.


Assuntos
Sistemas CRISPR-Cas , Neuroglia/efeitos dos fármacos , PTEN Fosfo-Hidrolase/genética , Proteínas Proto-Oncogênicas c-akt/genética , RNA Mensageiro/genética , Proteínas Supressoras de Tumor/genética , Processamento Alternativo , Sequência de Bases , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas , Meios de Cultivo Condicionados/química , Meios de Cultivo Condicionados/farmacologia , Edição de Genes , Engenharia Genética , Terapia Genética/métodos , Humanos , Neuroglia/metabolismo , Neuroglia/patologia , PTEN Fosfo-Hidrolase/metabolismo , PTEN Fosfo-Hidrolase/farmacologia , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Guia de Cinetoplastídeos/genética , RNA Guia de Cinetoplastídeos/metabolismo , RNA Mensageiro/metabolismo , Proteínas Supressoras de Tumor/metabolismo , Proteínas Supressoras de Tumor/farmacologia
16.
BMC Genomics ; 18(1): 129, 2017 02 06.
Artigo em Inglês | MEDLINE | ID: mdl-28166742

RESUMO

BACKGROUND: Partial pollen and embryo sac sterilities are the two main reasons for low fertility in autotetraploid rice. Our previous study revealed that small RNAs changes may associate with pollen fertility in autotetraploid rice. However, knowledge on comparative analysis between the development of pollen and embryo sac by small RNAs in autotetraploid rice is still unknown. In the present study, WE-CLSM (whole-mount eosin B-staining confocal laser scanning microscopy) and high-throughput sequencing technology was employed to examine the cytological variations and to analyze small RNAs changes during pollen and embryo sac development in autotetraploid rice compared with its diploid counterpart. RESULTS: A total of 321 and 368 differentially expressed miRNAs (DEM) were detected during pollen and embryo sac development in autotetraploid rice, respectively. Gene Ontology enrichment analysis on the targets of DEM associated with embryo sac and pollen development revealed 30 prominent functional gene classes, such as cell differentiation and signal transduction during embryo sac development, while only 7 prominent functional gene classes, such as flower development and transcription factor activity, were detected during pollen development in autotetraploid rice. The expression levels of 39 DEM, which revealed interaction with meiosis-related genes, showed opposite expression patterns during pollen and embryo sac development. Of these DEM, osa-miR1436_L + 3_1ss5CT and osa-miR167h-3p were associated with the female meiosis, while osa-miR159a.1 and osa-MIR159a-p5 were related with the male meiosis. 21 nt-phasiRNAs were detected during both pollen and embryo sac development, while 24 nt-phasiRNAs were found only in pollen development, which displayed down-regulation in autotetraploid compared to diploid rice and their spatial-temporal expression patterns were similar to osa-miR2275d. 24 nt TEs-siRNAs were found to be up-regulated in embryo sac but down-regulated in pollen development. CONCLUSION: The above results not only provide the small RNAs changes during four landmark stages of pollen and embryo sac development in autotetraploid rice but also have identified specifically expressed miRNAs, especially meiosis-related miRNAs, pollen-specific-24 nt-phasiRNAs and TEs-siRNAs in autotetraploid rice. Together, these findings provide a foundation for understanding the effect of polyploidy on small RNAs expression patterns during pollen and embryo sac development that may lead to different abnormalities in autotetraploid rice.


Assuntos
Perfilação da Expressão Gênica , Oryza/crescimento & desenvolvimento , Oryza/genética , Pólen/crescimento & desenvolvimento , Pequeno RNA não Traduzido/genética , Sementes/crescimento & desenvolvimento , Tetraploidia , Meiose/genética , MicroRNAs/genética , Oryza/citologia , Pólen/genética , RNA Interferente Pequeno/genética , Sementes/genética
17.
Lab Invest ; 96(8): 820-9, 2016 08.
Artigo em Inglês | MEDLINE | ID: mdl-27295346

RESUMO

Interleukin (IL)-33 is a novel IL-1 family member, and its administration has been associated with promotion of T helper type-2 (Th2) cell activity and cytokines, particularly IL-4 and IL-5 in vivo. Recently, IL-33 was shown to increase CD4(+)Foxp3(+) regulatory T cells (Tregs) and to suppress levels of the Th1-type cytokine IFN-γ in allogeneic heart transplantation in mice. Therefore, we hypothesized that IL-33 and leflunomide (Lef) could prolong graft survival in the concordant mouse-to-rat heart transplantation model. In this model, xenografts undergo acute humoral xenograft rejection (AHXR) typically on day 3 or cell-mediated rejection approximately on day 7 if AHXR is inhibited by Lef treatment. Recipients were treated with Lef (n=6), IL-33 (n=6), IL-33 combined with Lef (n=6), or left untreated (n=6) for survival studies. Heart grafts were monitored until they stopped beating. Mouse heterotopic grafts were performed, and recipients were sacrificed on days 2 and 7 for histological and flow cytometric analyses. The combination of IL-33 and Lef significantly prolonged the grafts from 17.3±2.3 to 2.8±0.4 days, compared to untreated controls. IL-33 administration with Lef, while facilitating Th2-associated cytokines (IL-4 on day 2 but not day 7), also decreased IFN-γ on day 2 and day 7, compared with Lef treatment only. Furthermore, IL-33 with Lef administration caused an expansion of suppressive CD4(+)Foxp3(+) Tregs in rats. The IL-33 and Lef combination therapy resulted in significantly prolonged graft survival, associated with markedly decreased Th1 cells and increased IL-10 levels. In addition, the combination therapy significantly decreased the percentage of CD-45(+) B cells on days 2 and 7, compared with monotherapy. These findings reveal a new immunoregulatory property of IL-33. Specifically, it facilitates regulatory cells, particularly functional CD4(+)Foxp3(+) Tregs that underlie IL-33-mediated cardiac xenograft survival. Moreover, it can decrease Th1 cells and cytokine expression of Th1 T cells in xenograft recipients, for example IFN-γ.


Assuntos
Sobrevivência de Enxerto/efeitos dos fármacos , Transplante de Coração , Imunossupressores/administração & dosagem , Interleucina-33/administração & dosagem , Isoxazóis/administração & dosagem , Animais , Linfócitos B/efeitos dos fármacos , Linfócitos B/imunologia , Citocinas/metabolismo , Sinergismo Farmacológico , Feminino , Fatores de Transcrição Forkhead/metabolismo , Sobrevivência de Enxerto/imunologia , Xenoenxertos , Imunidade Humoral/efeitos dos fármacos , Interferon gama/metabolismo , Leflunomida , Camundongos , Camundongos Endogâmicos C57BL , Ratos , Ratos Endogâmicos Lew , Proteínas Recombinantes/administração & dosagem , Baço/efeitos dos fármacos , Baço/imunologia , Linfócitos T Reguladores/efeitos dos fármacos , Linfócitos T Reguladores/imunologia , Células Th1/efeitos dos fármacos , Células Th1/imunologia
18.
World J Urol ; 34(2): 281-9, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26119349

RESUMO

PURPOSE: To investigate the impact of metabolic syndrome (MetS) on benign prostatic hyperplasia (BPH), focusing on MetS and its relationship with prostate volume and prostate-specific antigen (PSA) in Chinese patients by performing a meta-analysis. METHODS: We systematically searched the PubMed, Embase, China National Knowledge Infrastructure, Wanfang, and VIP databases from inception to November 2014. All studies investigating the impact of MetS on prostate volume and PSA among BPH patients were included. Pooled mean difference (WMD) and 95% confidence interval (CI) were used to analyze the difference between patients with MetS and those without MetS. RESULTS: Sixteen studies enrolled 1895 BPH patients, of whom 2224 had MetS. Compared with those without MetS, BHP patients with MetS had significantly higher total prostate volume (WMD 10.15 ml; 95% CI 7.37-12.93) and serum PSA level (WMD 0.53 ng/ml; 95% CI 0.17-0.88), respectively. In addition, annual prostate growth rate in patients with MetS was higher (WMD 0.49 ml/year; 95% CI 0.24-0.73) than in those without MetS. CONCLUSIONS: This meta-analysis supports that the presence of MetS increases total prostate volume and annual prostate growth rate in Chinese BPH patients. Future studies are needed to explain the detailed underlying mechanisms.


Assuntos
Síndrome Metabólica/epidemiologia , Hiperplasia Prostática/epidemiologia , China/epidemiologia , Humanos , Incidência , Masculino , Síndrome Metabólica/complicações , Hiperplasia Prostática/etiologia , Fatores de Risco
19.
Guang Pu Xue Yu Guang Pu Fen Xi ; 36(9): 2911-8, 2016 Sep.
Artigo em Zh | MEDLINE | ID: mdl-30084625

RESUMO

Dust-fall distribution of vegetation leaves can indicate the degree of air pollution; therefore the analysis of spatial characteristics of urban vegetation dust-fall has important practical significance for making more effective air pollution control policy. Based on the data of weight of dust, spectral reflectance and leaf area of Euonymus japonicus, Sophora japonica, poplar and davidiana collected in the main area of Beijing city, we compared the curve of spectrum of four plants "dust leaves" to "clean leaves"; the correlation analysis between leaf spectral reflectance ratio (Dust/Clean) of narrow band and satellite band was processed with the weight of dust-fall respectively, with application of four plants leaf data. Then, we built the regression model of the satellite band reflectance and NDVI with dustfall weight respectively, and we used the best model to retrieve the dust-fall distribution of vegetation coverage area in Beijing city, furthermore, we obtained the dust distribution of the whole Beijing city through interpolation. Finally, we carried out the rationality verification of the result by the land cover and land use of the high dust region, as well as the average concentration of PM10. The results showed that, dust leaves had an obviously lower reflectance than clean leaves in 780~1 300 nm which belonged to near-infrared bands; therewas a higher correlation between narrow band reflectance and dust-fall weight in 520~620 and 1 390~1 600 nm, up to -0.626; the coefficients of determination (R2) of inversion models were respectively 0.446 and 0.465,which were constructed by green band and NDVI of Landsat8 with dust-fall weight. Using the model established with NDVI to retrieving the dust-fall distribution of Beijing city, the results demonstrate that the distribution of dust-fall is high in north and low in south, high in east and low in west, high in downtown and low in the suburbs. This study offers a low-cost and effective method for investigating dust-fall distribution in urban area, and provides data support to analysis sources of pollution for the environmental protection department.

20.
Guang Pu Xue Yu Guang Pu Fen Xi ; 36(5): 1478-82, 2016 May.
Artigo em Zh | MEDLINE | ID: mdl-30001040

RESUMO

The direct absorption and utilization of low-molecular weight organic nitrogen (N) by soil microbial is a new subject in the research of microbial N nutrition. The study used gas chromatography-mass spectrometry to trace dual-labeled (13C, 15N) glycine from the soil solution and microorganisms. The results showed that glycine added to the soil was quickly taken up by soil microorganisms, with the half-life of glycine being 2.9 h. Withthe incubation of 4 h, the maximum amount of dual-labeled glycine in the microbial biomass was measured (equivalent to 10% of glycine added), indicating that added glycine was absorbed as intact molecular by soil microorganisms. The single labeled-Keto acid was detected in soil solution and in the microorganisms (decomposed production by double labeled glycine), but the content is extremely low, suggesting that added glycine mainly served as carbon (C) source for soil microbial life activities. This study demonstrated that compound specific stable dual labeled isotope analysis combined with chloroform fumigation technique was an effective method for detecting the low-molecular organic N utilized by soil microorganisms.


Assuntos
Microbiologia do Solo , Solo , Biomassa , Isótopos de Carbono , Cromatografia Gasosa-Espectrometria de Massas , Glicina , Marcação por Isótopo , Nitrogênio
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