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1.
J Biomed Nanotechnol ; 18(1): 77-86, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-35180901

RESUMO

An urgent problem in nursing care is the rising threat of cutaneous wound infections caused by harmful bacteria. In this study, we fabricated a series of cellulose acetate-hyaluronic acid (CA/HA) electrospun fibers loaded with berberine (BBR) using the electrospinning method to determine their antimicrobial performance and potential in in vivo skin wound dressing applications. The BBR-loaded CA/HA electrospun fibers (CA/HA/BBR) were analyzed using scanning electron and Fourier transform infrared microscopies; moreover, their mechanical properties were examined. The analyses demonstrated an average fiber diameter of 502 ± 50 nm; the tensile strength, Young's modulus, and break elongation of CA/HA electrospun fibers were approximately 3.23 ± 0.08 MPa, 17.5 ± 0.03 MPa, and 28.4%, respectively, whereas these values for CA/HA/BBR electrospun fibers were 238 ± 39 nm and 2.99 ± 0.05 MPa, 12.3 ± 0.04 MPa, and 47.8%, respectively. Antimicrobial evaluation of the CA/HA/BBR electrospun fibers demonstrated that the dressings made from these fibers exhibited greater antimicrobial efficacy (>95%) against Staphylococcus aureus and Escherichia coli when compared to that made from CA/HA (>80%) electrospun fibers. In vitro experiments showed that BBR loaded CA/HA electrospun fiber scaffolds have highly enhanced cell viability (>99) and proliferation of L929 fibroblastic cells after 7 days of incubation. In addition, in vivo evaluations in rats showed that the as-fabricated CA/HA/BBR bandage decreased wound size; moreover, it had accelerated healing ability (>95%) and collagen development with increasing treatment duration. These results showed that the addition of BBR enhanced the bioactivity of the dressing without damaging its physical characteristics. Thus, nanostructured dressing made of CA/HA/BBR electrospun fibers has excellent potency for tissue repair in nursing care.


Assuntos
Berberina , Nanofibras , Animais , Antibacterianos/farmacologia , Bandagens , Celulose/análogos & derivados , Ácido Hialurônico , Ratos , Cicatrização
2.
Mol Cell Biochem ; 477(1): 1-13, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34532813

RESUMO

Circular RNAs (circRNAs) have been identified as important regulators in cancer progression. Nevertheless, little is known about the biological function of circ_0000376 in the progression of osteosarcoma (OS). Cell viability, colony formation ability, apoptosis, and motility were analyzed by Cell Counting Kit-8 assay, colony formation assay, flow cytometry, and transwell assays. Cellular glycolytic metabolism was analyzed using commercial kits. RT-qPCR and Western blot assay were performed to analyze RNA and protein expression in OS tissues and cells. Starbase software was used to establish circRNA-microRNA (miRNA)-messenger RNA linkage, and intermolecular interaction was verified by dual-luciferase reporter assay. Xenograft tumor assay was conducted to analyze the effects of Tanshinone I (Tan I) and circ_0000376 on xenograft tumor growth in vivo. Tan I treatment suppressed the viability, migration, invasion, and glycolysis and triggered the apoptosis of OS cells. Tan I treatment markedly down-regulated circ_0000376 expression in OS cells. The addition of circ_0000376 plasmid largely rescued the malignant behaviors of OS cells upon Tan I exposure. Circ_0000376 interacted with miR-432-5p in OS cells. Circ_0000376 overexpression-mediated protective effects in Tan I-induced OS cells were partly attenuated by the accumulation of miR-432-5p. miR-432-5p bound to the 3' untranslated region (3'UTR) of B-cell leukemia/lymphoma 2 (BCL2) in OS cells. miR-432-5p interference-induced effects in Tan I-treated OS cells were partly overturned by the silence of BCL2. Circ_0000376 can act as miR-432-5p sponge to up-regulate BCL2 expression in OS cells. Circ_0000376 silencing contributed to the anti-tumor effect of Tan I on the growth of xenograft tumors in vivo. Tan I exerted an anti-tumor role in OS progression by targeting circ_0000376/miR-432-5p/BCL2 axis.


Assuntos
Abietanos/farmacologia , Neoplasias Ósseas/metabolismo , MicroRNAs/metabolismo , Osteossarcoma/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , RNA Circular/metabolismo , RNA Neoplásico/metabolismo , Transdução de Sinais , Animais , Neoplasias Ósseas/tratamento farmacológico , Neoplasias Ósseas/genética , Neoplasias Ósseas/patologia , Linhagem Celular Tumoral , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , MicroRNAs/genética , Osteossarcoma/tratamento farmacológico , Osteossarcoma/genética , Osteossarcoma/patologia , Proteínas Proto-Oncogênicas c-bcl-2/genética , RNA Circular/genética , RNA Neoplásico/genética , Ensaios Antitumorais Modelo de Xenoenxerto
3.
Aging (Albany NY) ; 13(9): 13333-13348, 2021 05 12.
Artigo em Inglês | MEDLINE | ID: mdl-33982674

RESUMO

This study aimed to explore effects of Sevoflurane on ischemia-reperfusion (I/R) injury after total knee arthroplasty (TKA). To explore potential molecular mechanism, Ras related dexamethasone induced 1 (RASD1), a Protein kinase A (PKA) activator, frequently associated with various models of I/R injury, was also investigated. In vivo mouse models with I/R injury after TKA and in vitro cell models with I/R injury were induced. Contents of creatinine kinase (CK), lactic dehydrogenase (LDH), superoxide dismutase (SOD), and malondialdehyde (MDA), serum levels of inflammatory factors, expression of PKA pathway-related genes and cell proliferation and apoptosis were measured. RASD1 was altered and PKA pathway was inhibited in mice and cells to elucidate the involvement of RASD1 and PKA pathway in Sevoflurane treatment on I/R injury. RASD1 was upregulated in I/R injury after TKA. Sevoflurane treatment or silencing RASD1 reduced RASD1 expression, CK, LDH and MDA contents, inflammation, apoptosis, but increased proliferation, SOD content, cAMP expression, and extents of PKA and cAMP responsive element binding protein (CREB) phosphorylation in skeletal muscle cells of I/R injury. Additionally, PKA pathway activation potentiated the therapeutic effect of Sevoflurane on I/R injury after TKA. Altogether, Sevoflurane treatment confines I/R injury after TKA via RASD1-mediated PKA pathway activation.


Assuntos
Proliferação de Células/efeitos dos fármacos , Traumatismo por Reperfusão/tratamento farmacológico , Sevoflurano/farmacologia , Proteínas ras/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Artroplastia do Joelho/métodos , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/genética , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Modelos Animais de Doenças , Camundongos , Substâncias Protetoras/farmacologia , Traumatismo por Reperfusão/metabolismo
4.
Comput Math Methods Med ; 2020: 7067649, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32855654

RESUMO

Osteosarcoma (OS) is one of the most common types of primary bone tumors in early adolescence with unsatisfied prognosis. Aberrant DNA methylation had been demonstrated to be related to tumorigenesis and progression of multiple cancers and could serve as the potential biomarkers for the prognosis of human cancers. In conclusion, this study identified 18 downregulated hypomethylation genes and 52 upregulated hypomethylation genes in OS by integrating the analysis the GSE97529 and GSE42572 datasets. Bioinformatics analysis revealed that OS-specific methylated genes were involved in regulating multiple biological processes, including chemical synaptic transmission, transcription, response to drug, and regulating immune response. KEGG pathway analysis showed that OS-specific methylated genes were associated with the regulation of Hippo, cAMP calcium, MAPK, and Wnt signaling pathways. By analyzing R2 datasets, this study showed that the dysregulation of these OS-specific methylated genes was associated with the metastasis-free survival time in patients with OS, including CBLN4, ANKMY1, BZW1, KRTCAP3, GZMB, KRTDAP, LY9, PFKFB2, PTPN22, and CLDN7. This study provided a better understanding of the molecular mechanisms underlying the progression and OS and novel biomarkers for the prognosis of OS.


Assuntos
Neoplasias Ósseas/genética , Metilação de DNA , DNA de Neoplasias/genética , Osteossarcoma/genética , Adolescente , Neoplasias Ósseas/metabolismo , Biologia Computacional , DNA de Neoplasias/metabolismo , Bases de Dados de Ácidos Nucleicos , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Redes Reguladoras de Genes , Humanos , Conceitos Matemáticos , Osteossarcoma/metabolismo , Prognóstico , Mapas de Interação de Proteínas/genética , Sarcoma de Ewing/genética , Sarcoma de Ewing/metabolismo , Sarcoma Sinovial/genética , Sarcoma Sinovial/metabolismo
5.
Biomed Pharmacother ; 129: 110239, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32590190

RESUMO

More and more researches have proved that long noncoding RNAs (lncRNAs) are vital regulators and biological participants in human cancers [1-5]. SnoRNA host gene 6 (SNHG6) was found to have an effect on the early stage and tumorigenesis in many cancers [6-10]. However, the expression of SNHG6 and its role of in non-small cell lung cancer (NSCLC) still need to be investigated. This work aims to investigate the expression and its biological role in NSCLC. In our study, the expression of SNHG6 was abnormally high in NSCLC tissues and cells. The negative impact of SNHG6 expression on the overall survival of patients with NSCLC was analyzed with Kaplan Meier method. Functionally, loss of SNHG6 expression led to the inhibition on the growth, migration and invasion of NSCLC cells. Mechanistically, miR-485-3p was necessary for the regulatory relation between SNHG6 and VPS45. More importantly, SPI1 could promote the expression of SNHG6 via transcriptionally activation. In conclusion, we proved that SPI1/SNHG6/miR-485-3p/VPS45 axis exerted oncogenic role in the cellular process of NSCLC cells.


Assuntos
Carcinoma Pulmonar de Células não Pequenas/metabolismo , Neoplasias Pulmonares/metabolismo , MicroRNAs/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , RNA Longo não Codificante/metabolismo , Transativadores/metabolismo , Proteínas de Transporte Vesicular/metabolismo , Apoptose , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , MicroRNAs/genética , Invasividade Neoplásica , Proteínas Proto-Oncogênicas/genética , RNA Longo não Codificante/genética , Transdução de Sinais , Transativadores/genética , Regulação para Cima , Proteínas de Transporte Vesicular/genética
6.
J Biochem Mol Toxicol ; 34(8): e22511, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32267991

RESUMO

To investigate the effect of microRNA-487b (miR-487b) as well as the underlying mechanism in osteosarcoma (OS). Data downloaded from the Gene Expression Omnibus (GEO) database were used to analyze the expression and prognostic value of miR-487b/TRAK2. Cell counting kit-8, colony formation, and transwell assays were performed to investigate the biological functions of miR-487b and TRAK2. Luciferase reporter assay was applied to confirm the interactions between miR-487b and TRAK2. miR-487b was overexpressed in OS tissues and was inversely associated with the prognosis of OS patients. We discovered that miR-487b could contribute to the proliferative, clonogenic, invasive, and migratory capabilities of OS cells. Through target prediction using miRWalk and differential expression analysis based on the GEO data set, trafficking kinesin protein 2 (TRAK2) was recognized as a potential target of miR-487b, which was further verified by luciferase reporter assay. The expression of TRAK2 was decreased in OS tissues compared with normal tissues and was positively correlated with the prognosis of OS patients. A negative relevance was presented between the expression of miR-487b and TRAK2 in OS cells. Of note, further mechanistic analyses indicated that TRAK2 was implicated in the regulatory effect of miR-487b on the cell malignant behaviors in OS. To sum up, these results demonstrated that miR-487b played an oncogenic role in OS progression via directly targeting TRAK2, which could advance the development of cancer treatment.


Assuntos
Biomarcadores Tumorais/biossíntese , Neoplasias Ósseas/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/biossíntese , MicroRNAs/biossíntese , Proteínas de Neoplasias/biossíntese , Proteínas do Tecido Nervoso/biossíntese , Osteossarcoma/metabolismo , RNA Neoplásico/biossíntese , Neoplasias Ósseas/diagnóstico , Neoplasias Ósseas/patologia , Linhagem Celular Tumoral , Humanos , Osteossarcoma/diagnóstico , Osteossarcoma/patologia , Prognóstico
7.
PLoS One ; 15(2): e0228964, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32049997

RESUMO

Jiaji Duck (JJ) is a Muscovy duck species that possesses many superior characteristics, and it has become an important genetic resource in China. However, to date, its genetic characteristics and genetic relationship with other duck breeds have not been explored yet, which greatly limits the utilization of JJ. In the present study, we investigated the genome sequences of 15 individual ducks representing five different duck populations, including JJ, French Muscovy duck (FF), mallard (YD), hong duck (HD) and Beijing duck (BD). Moreover, we investigated the characteristics of JJ-specific single nucleotide polymorphisms (SNPs) and compared the genome sequences of JJ vs. YD and JJ vs. BD using integrated strategies, including mutation detection, selective screening, and Gene Ontology (GO) analysis. More than 40 Gb of clean data were obtained for each population (mean coverage of 13.46 Gb per individual). A total number of 22,481,367 SNPs and 4,156,829 small insertion-deletions (Indels) were identified for the five duck populations, which could be used as molecular markers in breeding and utilization of JJ. Moreover, we identified 1,447,932 JJ-specific SNPs, and found that genes covering at least one JJ-specific SNP mainly involved in protein phosphorylation and dephosphorylation, as well as DNA modification. Phylogenetic tree and principal components analysis (PCA) revealed that the genetic relationship of JJ was closest to FF, while it was farthest to BD. A total of 120 and 111 genes were identified as positive selection genes for JJ vs. BD and JJ vs. YD, respectively. GO and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses showed that the positive selection genes for JJ vs. BD ducks mainly involved in pigmentation, muscle contraction and stretch, gland secretion, and immunology, while the positive selection genes obtained from JJ vs. YD ducks mainly involved in embryo development, muscle contraction and stretch, and gland secretion. Taken together, our findings enabled us to better understand the characteristics of JJ and provided a molecular basis for the breeding and hybrid utilization of JJ in the future.


Assuntos
Patos/genética , Genoma/genética , Animais , Cruzamento/métodos , China , Mapeamento Cromossômico/métodos , Feminino , Ontologia Genética , Mutação/genética , Fosforilação/genética , Filogenia , Polimorfismo de Nucleotídeo Único/genética , Análise de Componente Principal/métodos , Sequenciamento Completo do Genoma/métodos
8.
Cancer Cell Int ; 20: 2, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31911754

RESUMO

BACKGROUND: Lung adenocarcinoma (LUAD), a widespread histopathological subtype of lung cancer, is deemed as a malignant tumor with a peak risk of mortality. Emerged as RNA with a loop structure that depleted protein coding ability, circular RNA (circRNA) has been identified as a regulator in cancer progression. Circ-SOX4, identified as a novel circRNA, has not been studied in any cancer yet. Thus, the regulatory function that circ-SOX4 exerts on LUAD development remains obscure. AIM OF THE STUDY: This study aimed to investigate the biological function and molecular mechanism of circ-SOX4 in LUAD. METHODS: The expression of circ-SOX4 was detected by qRT-PCR. CCK-8, colony formation, transwell and wound healing assays were performed to explore the biological function of circ-SOX4 in LUAD. The interaction between miR-1270 and circ-SOX41 (or PLAGL2) was confirmed by RNA pull down, luciferase reporter and RIP assays. RESULTS: Circ-SOX4 was found to be obviously upregulated in LUAD tissues and cells, and knockdown of it inhibited cell proliferation, invasion and migration in LUAD. Furthermore, silenced circ-SOX4 also inhibited LUAD tumor growth. Molecular mechanism assays revealed that circ-SOX4 interacted with miR-1270 in LUAD. Besides, PLAGL2 was confirmed as a downstream gene of miR-1270. Rescue assays validated that miR-1270 suppression or PLAGL2 overexpression countervailed circ-SOX4 depletion-mediated inhibition on cell proliferation, invasion and migration in LUAD. Additionally, it was discovered that circ-SOX4/miR-1270/PLAGL2 axis activated WNT signaling pathway in LUAD. CONCLUSIONS: Circ-SOX4 boosted the development of LUAD and activate WNT signaling pathway through sponging miR-1270 and modulating PLAGL2, which provided a valuable theoretical basis for exploring underlying therapeutic target in LUAD.

9.
Biomed Res Int ; 2019: 3467121, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31737660

RESUMO

Spinal cord injury (SCI) is a highly debilitating disease and is increasingly being recognized as an important global health priority. However, the mechanisms underlying SCI have not yet been fully elucidated, and effective therapies for SCI are lacking. Long noncoding RNAs (lncRNAs), which form a major class of noncoding RNAs, have emerged as novel targets for regulating several physiological functions and mediating numerous neurological diseases. Notably, gene expression profile analyses have demonstrated aberrant changes in lncRNA expression in rats or mice after traumatic or nontraumatic SCI. LncRNAs have been shown to be associated with multiple pathophysiological processes following SCI including inflammation, neural apoptosis, and oxidative stress. They also play a crucial role in the complications associated with SCI, such as neuropathic pain. At the same time, some lncRNAs have been found to be therapeutic targets for neural stem cell transplantation and hydrogen sulfide treatment aimed at alleviating SCI. Therefore, lncRNAs could be promising biomarkers for the diagnosis, treatment, and prognosis of SCI. However, further researches are required to clarify the therapeutic effects of lncRNAs on SCI and the mechanisms underlying these effects. In this study, we reviewed the current progress of the studies on the involvement of lncRNAs in SCI, with the aim of drawing attention towards their roles in this debilitating condition.


Assuntos
Inflamação/genética , Neuralgia/genética , RNA Longo não Codificante/genética , Traumatismos da Medula Espinal/genética , Apoptose/genética , Regulação da Expressão Gênica , Humanos , Inflamação/complicações , Inflamação/fisiopatologia , Neuralgia/complicações , Neuralgia/fisiopatologia , Estresse Oxidativo/genética , Traumatismos da Medula Espinal/complicações , Traumatismos da Medula Espinal/fisiopatologia , Transcriptoma
10.
Virol Sin ; 32(3): 199-206, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28530022

RESUMO

The Zika virus (ZIKV) is an arbovirus that has spread rapidly worldwide within recent times. There is accumulating evidence that associates ZIKV infections with Guillain-Barré Syndrome (GBS) and microcephaly in humans. The ZIKV is genetically diverse and can be separated into Asian and African lineages. A rapid, sensitive, and specific assay is needed for the detection of ZIKV across various pandemic regions. So far, the available primers and probes do not cover the genetic diversity and geographic distribution of all ZIKV strains. To this end, we have developed a one-step quantitative reverse transcription polymerase chain reaction (qRT-PCR) assay based on conserved sequences in the ZIKV envelope (E) gene. The detection limit of the assay was determined to be five RNA transcript copies and 2.94 × 10-3 50% tissue culture infectious doses (TCID50) of live ZIKV per reaction. The assay was highly specific and able to detect five different ZIKV strains covering the Asian and African lineages without nonspecific amplification, when tested against other flaviviruses. The assay was also successful in testing for ZIKV in clinical samples. Our assay represents an improvement over the current methods available for the detection ZIKV and would be valuable as a diagnostic tool in various pandemic regions.


Assuntos
Técnicas de Diagnóstico Molecular/métodos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Infecção por Zika virus/diagnóstico , Zika virus/isolamento & purificação , Sequência Conservada , Primers do DNA/genética , Humanos , Sensibilidade e Especificidade , Proteínas do Envelope Viral/genética , Zika virus/genética , Infecção por Zika virus/virologia
11.
Mol Biol Rep ; 39(7): 7647-55, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22451153

RESUMO

Pekin Duck is world-famous for its fast growth, but its breast muscle development is later and breast muscle content is lower compared with other muscular ducks. Therefore, it is very important to discover the genetic mechanism between breast muscle development and relative gene expression in Pekin duck. In current study, the genes which have relationships with breast muscle development were identified by suppression subtractive hybridization. A total of 403 positive clones were sequenced and 257 unigenes were obtained. The expression of 23 genes were analyzed in the breast muscle of 2-, 4-, 6-, 8- week old Pekin ducks. The results showed that unknown clone A233, C83 and C99 showed descending tendency as age increased; KBTBD10, HSPA8, MYL1, ZFP622, MARCH4, Nexilin, FABP4 and MUSTN1 had high expression levels at 6 weeks old; WAC, NT5C3, HSP90AA1, MRPL33, KLF6, TSNAX, CDC42EP3, HSPA4, TRAK1, NR2F2, HAUS1 and IGF1 had high expression levels at 8 weeks and showed ascending tendency as age increased. Expression of these 23 genes were also analyzed in breast muscle, leg muscle, heart, kidney, liver, muscular stomach and sebum cutaneum in 4-8-week old Pekin duck and results showed that most of these genes had high expression in breast muscle, leg muscle and heart.


Assuntos
Patos/genética , Patos/fisiologia , Desenvolvimento Muscular/genética , Músculo Esquelético/crescimento & desenvolvimento , Animais , Sequência de Bases , Patos/crescimento & desenvolvimento , Feminino , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Genes , Masculino , Dados de Sequência Molecular , Análise de Sequência de DNA , Tórax/crescimento & desenvolvimento
12.
Zhongguo Wei Zhong Bing Ji Jiu Yi Xue ; 22(9): 547-52, 2010 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-20854736

RESUMO

OBJECTIVE: To investigate the modulation effect of ulinastatin (UTI) preconditioning on gene expression of kidney tissue in septic rats by DNA microarrays. METHODS: Forty-five male Wistar rats were divided into control group, sepsis group and UTI group, with 15 rats in each group by means of random number table. Cecal ligation and puncture (CLP) was used to reproduce rat sepsis model. The control group only experienced a simulated operation without CLP. In UTI group the rats were treated with intramuscular injection of UTI (100 kU/kg). In sepsis group and control group intramuscular balanced solution (5 ml/kg) was given. Gene expression spectrum was studied with oligonucleotide gene expression profile microarray that contained 22 523 rat cDNA clones to detect the changes in gene expression pattern of rat kidney tissue 24 hours after CLP. Genes with fluorescent signal of Cy3/Cy5 of ratio average (RA)>2.0 or RA<0.5 were identified as differential genes, then those highly correlated to sepsis and UTI were screened by means of related computer software, and their relationship was analyzed. RESULTS: Three hundred and twenty-seven differential genes were found in sepsis group/control group, accounting for 1.45%, and among them 181 genes showed up-regulation,with 78 known functional genes, and 146 genes showed down-regulation, with 51 known functional genes. One hundred and twenty-seven differential genes were found in UTI group/sepsis group, accounting for 0.56%, and among them 41 genes showed up-regulation, with 14 known functional genes, and 86 genes showed down-regulation, with 37 known functional genes. Twenty-two genes were down-regulated in sepsis group/control group but up-regulated in UTI group/sepsis group, with 11 known functional genes, 51 genes were up-regulated in sepsis group/control group but down-regulated in UTI group/sepsis group, with 24 known functional genes. CONCLUSION: UTI preconditioning can alleviate the damage of kidney tissue in rat sepsis model, thus showing a protective effect on kidney, and the mechanism may be attributable to effect of UTI on modulation of immune reaction, energy metabolism, inflammatory reaction, signal transduction, defense reaction, oxidation-reduction reaction, DNA replication, and transcription related genes.


Assuntos
Glicoproteínas/farmacologia , Rim/metabolismo , Sepse/metabolismo , Transcriptoma , Animais , Modelos Animais de Doenças , Perfilação da Expressão Gênica , Inflamação , Masculino , Análise de Sequência com Séries de Oligonucleotídeos , Ratos , Ratos Wistar
13.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 18(3): 638-41, 2010 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-20561418

RESUMO

This study was aimed to investigate the effects of sodium valproate (VPA) on the proliferation and regulation of histone acetylation of multiple myeloma cell line U266. U266 cells were treated with VPA. Cell proliferation was determined by CCK-8 assay, and cell cycle were analyzed by flow cytometry (FCM). The expression level of HDAC1 mRNA was detected by RT-PCR, and the protein levels of HDAC1 and histone H3, H4 acetylation was detected by Western blot. The results showed that the VPA inhibited the proliferation of U266 cells in concentration-and time-dependent manners.After exposure to different concentrations of VPA for 48 hours, the proportion of G(0)/G(1) cells increased, while the proportion of S phase cells decreased. The cell cycle was arrested obviously in G(0)/G(1) phase (p < 0.05). The expression of HDAC1 mRNA was inhibited, and the protein level of HDAC1 was down-regulated, while the histone H3/H4 acetylation was up-regulated in U266 cells. It is concluded that the VPA can inhibit cell proliferation of U266 and induce G(0)/G(1) phase arrest. The increase of histone H3/H4 acetylation resulting from inhibiting HDAC1 by VPA might be considered as a possible mechanism.


Assuntos
Proliferação de Células/efeitos dos fármacos , Inibidores de Histona Desacetilases/farmacologia , Histonas/metabolismo , Mieloma Múltiplo/metabolismo , Ácido Valproico/farmacologia , Acetilação/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Histona Desacetilase 1/metabolismo , Humanos
14.
Zhonghua Xue Ye Xue Za Zhi ; 31(12): 835-8, 2010 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-21223742

RESUMO

OBJECTIVE: To study the antitumour effects of sodium valproate (VPA) on the proliferation, differentiation and cell cycle of Molt-4 cell and to investigate its demethylation mechanisms. METHODS: After Molt-4 cells trated with VPA at different concentrations, cell viability and growth curve were assessed by MTT assay. Cell cycle changes were analyzed by flow cytometry. The expression level of p15, DNA methyltransferase 1 (DNMT-1), DNMT3A and 3B mRNA were detected by RT-PCR and the methylation level was detected by hn-MSPCR. RESULTS: VPA significantly inhibited the proliferation of Molt-4 cells. After 48 h culture with 5.0 mmol/L VPA, the percentages of Molt-4 cells in G(0)/G(1) phase was (66.87 ± 3.31)% and in S phase was (8.47 ± 2.56)%, while in control group, the cells in G(0)/G(1) phase increased and in S phase decreased significantly. The p15 gene in Molt-4 cells failed to express due to its hypermethylation. The expression level of p15 gene mRNA increased significantly after exposure to VPA for 48 h. As compared with control group, the expression of DNMT-1 was down-regulated in a dose-dependent manner. The expression level of DNMT3B decreased at 10.0 mmol/L concentration. CONCLUSION: VPA has a demethylation effect on p15 INK4B gene by inhibiting the DNMT-1 and DNMT3B gene activities to recover p15 gene activity, which arrests Molt-4 cell in G(0)/G(1) phase.


Assuntos
Metilação de DNA , Ácido Valproico , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Metilação de DNA/efeitos dos fármacos , RNA Mensageiro/genética , Ácido Valproico/farmacologia
15.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 16(6): 1288-92, 2008 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-19099629

RESUMO

This study was purposed to investigate the synergistic effects of sodium valproate (VPA) and As2O3 on the proliferation of Molt-4 cells in vitro and its possible mechanisms. Cell viability and growth curve were assessed by the MTT assay. The synergistic activity in combination of 2 drugs was determined by the Q format. The expression levels of p15, DNA methyltransferase 1 (DNMT-1), DNMT3A and DNMT 3B mRNA were detected by RT-PCR and the methylation level was detected by hn-MSPCR. The results indicated that the VPA and As2O3 both inhibited proliferation of Molt-4 cells. The combination of two drugs showed an additive effect (values of Q were>or=0.85). The inhibitory rate in combination of 5 mmol/L of VPA with 10 micromol/L of As2O3 was (70.31+/-2.54)%. The p15 gene in Molt-4 cell line failed to express due to its hypermethylation. The level of p15 gene mRNA expression increased significantly after exposure to VPA in combination with As2O3 for 48 h. As compared with control group, the expression of DNMT-1 was down-regulated in a dose-dependent manner, whereas DNMT3A had no significant differences from the control. The level of expression of DNMT3B seemed to decrease at 10 mmol/L concentration. There were significant differences between the combination of the two drugs and the control group. The gray value of methylated bands decreased after the treatment of VPA alone and in combination with As2O3 in a dose-dependent manner. It is concluded that VPA induces demethylation of p15 INK4B gene by inhibiting the DNMT-1 and DNMT3B gene activities, which up-regulates the p15 gene, recovers its activity. The combination of VPA with As2O3 has the synergistic additive effect on the inhibition of cell viability, so that VPA can reduce the side effect of As2O3 on liver function, which would be verified in the clinical practice.


Assuntos
Arsenicais/farmacologia , Inibidor de Quinase Dependente de Ciclina p15/metabolismo , Óxidos/farmacologia , Ácido Valproico/farmacologia , Trióxido de Arsênio , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Inibidor de Quinase Dependente de Ciclina p15/genética , DNA (Citosina-5-)-Metiltransferase 1 , DNA (Citosina-5-)-Metiltransferases/metabolismo , Metilação de DNA/efeitos dos fármacos , DNA Metiltransferase 3A , Sinergismo Farmacológico , Regulação Neoplásica da Expressão Gênica , Humanos , Regulação para Cima
16.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 15(2): 258-61, 2007 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-17493327

RESUMO

The study was aimed to explore the relationship between patterns of methylation or deletion and the development of acute leukemia, and further to clarify the possible mechanism in the development of adult acute leukemia. Nested methylation-specific polymerase chain reaction (n-MSP) was adopted to analyze p16 gene methylation or deletion patterns in 82 adult acute leukemia patients with different subtypes and stages. The results indicated that rate of p16 gene methylation was 39.0% in 82 adult acute leukemia patients, among them, 41.4% in acute myelogenous leukemia (AML) and 33.3% in acute lymphoblastic leukemia (ALL). It were found that 36.6% of de novo AL patients and 54.5% of relapsed AL patients developed the hypermethylation of p16 gene. Out of the 82 patients, 6 seemed to have deletion of p16 gene, including 1 AML (1.7%) and 5 ALL (20.8%). There were no hypermethylation or deletion of p16 gene in the 16 controls. It is concluded that methylation of p16 gene may play a more important role than homozygous deletion of p16 gene in the leukemogenesis and progression of adult acute leukemia.


Assuntos
Ilhas de CpG/genética , Metilação de DNA , Genes p16 , Leucemia Mieloide Aguda/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Adolescente , Adulto , Idoso , Sequência de Bases , DNA de Neoplasias/genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/métodos
17.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 15(2): 382-6, 2007 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-17493352

RESUMO

This study was aimed to investigate the methylation or deletion status of p15 gene in different malignant cell lines, and further to clarify their roles in the development and progression of malignant tumors. Hemi-nested methylation specific polymerase chain reaction (hn-MSP) was adopted to analyze p15 gene methylation or deletion status in 20 malignant tumor cell lines and mononuclear cells or normal cell lines in healthy people, as well as to evaluate its sensitivity and specificity. The results showed that among all of the cell lines, Molt-4, KG1, NCE, Raji, SMMC-7221, CA46, SW480 and NCI-H446 were partial methylated with CDKN2B gene, and its sensitivity of detection of p15 gene methylation was up to 1.0 x 10(-5), also it had great specificity. Peripheral blood mononuclear cell (MNCs) from healthy volunteer, HL-60, HepG2, 293, HeLa, SGC7901, U266 and CEM were unmethylated; and K562, NB4, GMC, Jurkat seemed to have deletion or mutation of p15 gene. It is concluded that the incidence of p15 gene methylation or deletion in many tumours, especially malignant hematopathy, is frequent, they correlate with disease progression and prognosis. Hn-MSP is highly sensitive and specific in analyzing p15 gene methylation, deserving in clinical application.


Assuntos
Ilhas de CpG/genética , Inibidor de Quinase Dependente de Ciclina p15/genética , Metilação de DNA , Deleção de Genes , Neoplasias Hematológicas/genética , Regulação Neoplásica da Expressão Gênica , Neoplasias Hematológicas/patologia , Humanos , Reação em Cadeia da Polimerase/métodos , Células Tumorais Cultivadas
18.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 14(2): 375-8, 2006 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-16638219

RESUMO

This study was aimed to investigate the efficiency of modified methylation-specific polymerase chain reaction i.e. nested methylation-specific polymerase chain reaction, used to detect the promoter methylation of p16 gene in six hematological malignant cell lines, and to explore the application in selection of hematological malignant cell lines with promoter hypermethylation, and make them to be an idel cell models for studying the relationship between gene methylation and expression. DNAs were denatured by NaOH and then were subjected to bisulfite modification and a nested-MSP was used to amplify the promoter region, nested MSP product of p16 gene promoter was analyzed and sequenced. The results showed that the hypermethylation of p16 gene was detected in CA46 and U266, however, Molt4, K562, HL-60 and Jurkat cell lines were unmethylated. In conclusion, p16 gene methylation in hematological malignant cell lines can be perfectly detected by nested-MSP method, which is simple, sensitive and specific for screening all kinds of hematological malignant cell lines with p16 gene methylated.


Assuntos
Metilação de DNA , Genes p16 , Neoplasias Hematológicas/genética , Regiões Promotoras Genéticas/genética , Sequência de Bases , Linhagem Celular Tumoral , Células HL-60 , Neoplasias Hematológicas/patologia , Humanos , Células K562 , Linfoma/genética , Linfoma/patologia , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/métodos , Análise de Sequência de DNA
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